Details of experiments between P61978 and O00499
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: HNRNPK
Protein A fragment: 1-463
Protein A construct: endogenous protein (Jurkat)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.22
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
Measured pKd: 4.55


Experiment series 2

Protein A protein: HNRNPK
Protein A fragment: 1-463
Protein A construct: endogenous protein (Jurkat)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.35
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.77
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.4
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.6


Experiment series 4

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.22
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.21


Experiment series 5

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.3
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.41


Experiment series 6

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.43
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.67


Experiment series 7

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.43
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.66


Experiment series 8

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 3.9


Experiment series 9

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.38
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.56


Experiment series 10

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.17
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.07


Experiment series 11

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.25
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.3


Experiment series 12

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.35
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.51


Experiment series 13

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.37
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.55


Experiment series 14

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.1
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 3.79


Experiment series 15

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.07
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.65


Experiment series 16

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: -0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 17

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 18

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.06
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.57


Experiment series 19

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.06
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.53


Experiment series 20

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.02
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 21

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: -0
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 22

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.02
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 23

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 24

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: -0.07
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 25

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: -0.01
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 26

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: -0
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 27

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.18
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.59


Experiment series 28

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.08
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.19


Experiment series 29

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.1
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.29


Experiment series 30

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.15
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.51


Experiment series 31

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.14
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.47


Experiment series 32

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.05
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.96


Experiment series 33

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.4
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.6


Experiment series 34

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.21
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.18


Experiment series 35

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.24
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.27


Experiment series 36

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.34
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.48


Experiment series 37

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.3
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.4


Experiment series 38

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.13
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.94


Experiment series 39

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.33
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.47


Experiment series 40

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.15
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.01


Experiment series 41

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.19
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.13


Experiment series 42

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.32
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.46


Experiment series 43

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.31
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.42


Experiment series 44

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.06
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.53


Experiment series 45

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.35
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.51


Experiment series 46

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.17
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.06


Experiment series 47

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.19
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.13


Experiment series 48

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.34
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.49


Experiment series 49

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.33
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.47


Experiment series 50

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.07
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.65


Experiment series 51

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.24
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 9
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.58


Experiment series 52

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.09
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 9
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.08


Experiment series 53

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.1
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 9
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.12


Experiment series 54

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.21
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 9
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.52


Experiment series 55

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.18
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 9
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.43


Experiment series 56

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 9
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 57

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.37
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.54


Experiment series 58

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.18
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.09


Experiment series 59

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.2
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.16


Experiment series 60

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.34
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.48


Experiment series 61

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.35
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.52


Experiment series 62

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.09
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.73


Experiment series 63

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.04
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 64

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.03
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 65

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.06
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.52


Experiment series 66

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.05
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 67

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.03
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 68

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 69

Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.18
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.61


Experiment series 70

Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.09
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.22


Experiment series 71

Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.12
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.36


Experiment series 72

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.18
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.59


Experiment series 73

Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.16
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.53


Experiment series 74

Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 75

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.53
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.84


Experiment series 76

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.53
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.85


Experiment series 77

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.57
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 4.92


Experiment series 78

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 3.88


Experiment series 79

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 13
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 4.81


Experiment series 80

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.31
Immobilized partner concentration in holdup experiment (10-6M): 8
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 4.79


Experiment series 81

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 82

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.88


Experiment series 83

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.34
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 4.77


Experiment series 84

Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.54
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 4.87