Details of experiments between Q9UKJ3 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: GPATCH8
Protein A fragment: 1-1502
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: 0.34
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): 6.51
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: 4.72


Experiment series 2

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-CONH2

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

Average holdup BI: -0.04
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
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-CONH2

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

Average holdup BI: 0.84
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: 5.54


Experiment series 4

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: -0.03
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
The affinity was below the detection threshold of the assay.


Experiment series 5

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.84
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: 5.55


Experiment series 6

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: -0.03
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 7

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.78
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: 5.38


Experiment series 8

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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
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 9

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.48
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: 4.76


Experiment series 10

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: -0.02
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
The affinity was below the detection threshold of the assay.


Experiment series 11

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.71
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: 5.38


Experiment series 12

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: -0.01
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
The affinity was below the detection threshold of the assay.


Experiment series 13

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.65
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: 5.48


Experiment series 14

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: -0.07
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 15

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.1
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: 3.8


Experiment series 16

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: 0.01
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
The affinity was below the detection threshold of the assay.


Experiment series 17

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.53
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 4.86


Experiment series 18

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: -0.05
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
The affinity was below the detection threshold of the assay.


Experiment series 19

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.71
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: 5.52


Experiment series 20

Protein A protein: GPATCH8
Protein A fragment: 602-616
Protein A site: PxxP_1
Protein A sequence: HLQGLDPGEPNKSKE
Protein A construct: Bio Ado Ado-HLQGLDPGEPNKSKE-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: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 21

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.75
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: 5.3