Details of experiments between O75582 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: RPS6KA5
Protein A fragment: 1-802
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: 0.44
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): 5.13
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: 4.89
Experiment series 2
Protein A protein: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.03
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 3
Protein A protein: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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: -0.01
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: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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: -0.01
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 5
Protein A protein: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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: 0.08
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: 3.67
Experiment series 6
Protein A protein: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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 7
Protein A protein: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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 8
Protein A protein: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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
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: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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: -0.02
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: RPS6KA5
Protein A fragment: 270-284
Protein A site: PxxP_1
Protein A sequence: RILKSEPPYPQEMSA
Protein A construct: Bio Ado Ado-RILKSEPPYPQEMSA-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: 0.05
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: 3.47