Details of experiments between the fragments Q7Z6E9~725-739 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: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-CONH2

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

Average holdup BI: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.64


Experiment series 2

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.64


Experiment series 3

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.36
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: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 3.96


Experiment series 5

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.25
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: 4.44


Experiment series 6

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.25
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: 4.66


Experiment series 7

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.07
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: 3.61


Experiment series 8

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.32
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: 4.45


Experiment series 9

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.26
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: 4.6


Experiment series 10

Protein A protein: RBBP6
Protein A fragment: 725-739
Protein A site: PxxP_3
Protein A sequence: RSYSRSPPYPRRGRG
Protein A construct: Bio Ado Ado-RSYSRSPPYPRRGRG-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: 0.45
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: 4.69