Details of experiments between the fragments Q8WXE0~863-877 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: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.49
Holdup BI standard deviation: 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: 2
Measured pKd: 4.77
Experiment series 2
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.49
Holdup BI standard deviation: 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: 2
Measured pKd: 4.78
Experiment series 3
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.53
Holdup BI standard deviation: 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: 2
Measured pKd: 4.84
Experiment series 4
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.21
Holdup BI standard deviation: 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: 2
Measured pKd: 4.19
Experiment series 5
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.42
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 4.81
Experiment series 6
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.26
Holdup BI standard deviation: 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: 2
Measured pKd: 4.67
Experiment series 7
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: -0.02
Holdup BI standard deviation: 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.05
Holdup BI standard deviation: 0.02
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: 2
Measured pKd: 3.48
Experiment series 9
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.31
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 4.7
Experiment series 10
Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-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: 0.51
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 2
Measured pKd: 4.8