Details of experiments between the fragments O00499~513-593 and Q9NZQ3~238-252
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: his6-MBP-TEVsite-SH3
Protein B protein: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
Average holdup BI: 0.43
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: 4.67
Experiment series 2
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: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
Average holdup BI: 0.45
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: 4.71
Experiment series 3
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: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
Average holdup BI: 0.51
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: 4.82
Experiment series 4
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: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
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: 2023/01/17
Number of measurements: 1
Measured pKd: 3.98
Experiment series 5
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: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
Average holdup BI: 0.31
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: 4.57
Experiment series 6
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: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
Average holdup BI: 0.26
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: 4.68
Experiment series 7
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: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-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: 2023/01/17
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: Y531S
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
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
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: R581C
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
Average holdup BI: 0.23
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: 4.52
Experiment series 10
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: NCKIPSD
Protein B fragment: 238-252
Protein B site: PxxP_4
Protein B sequence: SCSPTPPPVPRRGTH
Protein B construct: Bio Ado Ado-SCSPTPPPVPRRGTH-CONH2
Average holdup BI: 0.49
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: 4.77