Details of experiments between Q8IY57 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: YAF2
Protein A fragment: 1-180
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: 0.59
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): 5.72
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: 5.15
Experiment series 2
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.49
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 2
Measured pKd: 4.78
Experiment series 3
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.51
Holdup BI standard deviation: 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: 2
Measured pKd: 4.81
Experiment series 4
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.51
Holdup BI standard deviation: 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: 2
Measured pKd: 4.81
Experiment series 5
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.04
Holdup BI standard deviation: 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.33
Holdup BI standard deviation: 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: 2
Measured pKd: 4.62
Experiment series 7
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.26
Holdup BI standard deviation: 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: 2
Measured pKd: 4.68
Experiment series 8
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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
Holdup BI standard deviation: 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.41
Holdup BI standard deviation: 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: 2
Measured pKd: 4.62
Experiment series 10
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.3
Holdup BI standard deviation: 0
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: 2
Measured pKd: 4.68
Experiment series 11
Protein A protein: YAF2
Protein A fragment: 3-17
Protein A site: PxxP_1
Protein A sequence: DKKSPTRPKRQPKPS
Protein A construct: Bio Ado Ado-DKKSPTRPKRQPKPS-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: 0.5
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 2
Measured pKd: 4.8