Details of experiments between the fragments Q96DX4~74-88 and O00499~513-593
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.79
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: 5.41
Experiment series 2
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.8
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: 5.43
Experiment series 3
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.75
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: 5.29
Experiment series 4
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.46
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: 4.73
Experiment series 5
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.66
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: 5.27
Experiment series 6
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.6
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: 5.36
Experiment series 7
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: -0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.71
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: 5.22
Experiment series 9
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.62
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: 5.34
Experiment series 10
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-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: 0.75
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: 5.29