Details of experiments between O00499 and Q96BZ8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

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

Protein B protein: LENG1
Protein B fragment: 1-264
Protein B construct: endogenous protein (Jurkat)

Average holdup BI: 0.49
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): 3.5
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: 4.98

Experiment series 2

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.

Experiment series 3

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.

Experiment series 4

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

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 5

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

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 6

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

Average holdup BI: -0.01
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 7

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

Average holdup BI: -0.02
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 8

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

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 9

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

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 10

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

Average holdup BI: -0.02
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 11

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

Protein B protein: LENG1
Protein B fragment: 184-198
Protein B site: PxxP_1
Protein B sequence: GSEKQRPKEPPSLDQ
Protein B construct: Bio Ado Ado-GSEKQRPKEPPSLDQ-CONH2

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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.