Details of experiments between the fragments Q8WXE0~785-799 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: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.34
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: 4.49
Experiment series 2
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.37
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: 4.55
Experiment series 3
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.42
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: 4.65
Experiment series 4
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.04
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 5
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.16
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: 4.18
Experiment series 6
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.13
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: 4.3
Experiment series 7
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: -0.03
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: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.17
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: 4.07
Experiment series 9
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.14
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: 4.24
Experiment series 10
Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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: 0.37
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: 4.56