Details of experiments between the fragments O43516~393-407 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: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.06
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: 3.52
Experiment series 2
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.08
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: 3.68
Experiment series 4
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.04
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
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.07
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: 4.01
Experiment series 7
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 3.5
Experiment series 8
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.06
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: 3.83
Experiment series 10
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-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: 0.08
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: 3.69