Details of experiments between O00499 and Q9NQZ6
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: ZC4H2
Protein B fragment: 1-224
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.48
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): 4.1
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: 4.97
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.29
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: 4.37
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.26
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: 4.32
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.31
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: 4.42
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
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 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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.15
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: 4.16
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.12
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: 4.24
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
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: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Y531S
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.44
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: 4.68
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.14
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: 4.25
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: ZC4H2
Protein B fragment: 209-223
Protein B site: PxxP_1
Protein B sequence: KSRSRNPKKPKRKQD
Protein B construct: Bio Ado Ado-KSRSRNPKKPKRKQD-CONH2
Average holdup BI: 0.31
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: 4.43