Details of experiments between the fragments O00499~513-593 and Q15287~252-266
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-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 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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.28
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: Q540H
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.36
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: 4.53
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.18
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: 4.11
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.27
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: 4.49
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.21
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: 4.56
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.19
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: 4.12
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-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: 2023/01/18
Number of measurements: 1
Measured pKd: 3.96
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.19
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: 4.41
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: RNPS1
Protein B fragment: 252-266
Protein B site: PxxP_2
Protein B sequence: PPRRMLPPPPMWRRS
Protein B construct: Bio Ado Ado-PPRRMLPPPPMWRRS-CONH2
Average holdup BI: 0.36
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: 4.53