Details of experiments between the fragments Q9UKJ3~735-749 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: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.84
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: 5.54


Experiment series 2

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.84
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: 5.55


Experiment series 3

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.78
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: 5.38


Experiment series 4

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.48
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: 4.76


Experiment series 5

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.71
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: 5.38


Experiment series 6

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.65
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: 5.48


Experiment series 7

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.1
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: 3.8


Experiment series 8

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.53
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: 4.86


Experiment series 9

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 0.71
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: 5.52


Experiment series 10

Protein A protein: GPATCH8
Protein A fragment: 735-749
Protein A site: PxxP_2
Protein A sequence: PPGSGSPAPPRRRRR
Protein A construct: Bio Ado Ado-PPGSGSPAPPRRRRR-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: 5.3