Details of experiments between the fragments O00499~513-593 and Q9Y6X9~725-739
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

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

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.29
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: 2023/01/16
Number of measurements: 2
Measured pKd: 4.38


Experiment series 2

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: T532M
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.29
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: 4.38


Experiment series 3

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Q540H
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.35
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: 4.51


Experiment series 4

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: D537V
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.01
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 5

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: V583I
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.16
Holdup BI standard deviation: 0
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: 4.19


Experiment series 6

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: P551L
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.12
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: 4.24


Experiment series 7

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: F584S
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: -0
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
The affinity was below the detection threshold of the assay.


Experiment series 8

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Y531S
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.03
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: 2023/01/18
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 9

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: R581C
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.14
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: 4.22


Experiment series 10

Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: V566M
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MORC2
Protein B fragment: 725-739
Protein B site: PxxP_3
Protein B sequence: SPIKLSPATPSRKRS
Protein B construct: Bio Ado Ado-SPIKLSPATPSRKRS-CONH2

Average holdup BI: 0.3
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: 4.4