Details of experiments between Q8IY67 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: RAVER1
Protein A fragment: 1-606
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: 0.63
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): 6.53
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: 5.23


Experiment series 2

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-CONH2

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

Average holdup BI: 0.56
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: 4.91


Experiment series 3

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-CONH2

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

Average holdup BI: 0.15
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: 4.02


Experiment series 4

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.57
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.92


Experiment series 5

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: 0.14
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: 3.98


Experiment series 6

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.6
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: 4.97


Experiment series 7

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: 0.18
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: 4.1


Experiment series 8

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.09
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: 3.77


Experiment series 9

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: 0.28
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: 4.36


Experiment series 10

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.4
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: 4.77


Experiment series 11

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: 0.06
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: 3.67


Experiment series 12

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.33
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: 4.84


Experiment series 13

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: 0.05
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: 3.85


Experiment series 14

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: -0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 15

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: -0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 16

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.24
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: 4.26


Experiment series 17

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: -0.02
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 18

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.37
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: 4.83


Experiment series 19

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: 0.04
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 20

Protein A protein: RAVER1
Protein A fragment: 420-434
Protein A site: PxxP_1
Protein A sequence: AGGGLPPELPPRRGK
Protein A construct: Bio Ado Ado-AGGGLPPELPPRRGK-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: 0.59
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: 4.96


Experiment series 21

Protein A protein: RAVER1
Protein A fragment: 431-445
Protein A site: PxxP_2
Protein A sequence: RRGKPPPLLPSVLGP
Protein A construct: Bio Ado Ado-RRGKPPPLLPSVLGP-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: 0.11
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: 3.83