Details of experiments between O00499 and A8CG34
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: avi(biotin)-his6-MBP-TEVsite-SH3

Protein B protein: POM121C
Protein B fragment: 1-1229
Protein B construct: endogenous protein (Jurkat)

Average holdup BI: 0.75
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.52
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
Measured pKd: 5.47


Experiment series 2

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

Protein B protein: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 3

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

Protein B protein: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: 0.19
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: 4.12


Experiment series 4

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

Protein B protein: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

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: 2023/01/16
Number of measurements: 1
Measured pKd: 3.96


Experiment series 5

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: -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: 1
The affinity was below the detection threshold of the assay.


Experiment series 6

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: 0.2
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: 4.15


Experiment series 7

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: 0.16
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: 4.04


Experiment series 8

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: -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: 1
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: Q540H
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: 0.23
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: 4.25


Experiment series 10

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: 0.16
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: 4.03


Experiment series 11

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: 0.02
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 12

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

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: 2023/01/17
Number of measurements: 1
Measured pKd: 3.53


Experiment series 13

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.


Experiment series 14

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: 0.01
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 15

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: 0.12
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: 4.05


Experiment series 16

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: 0.02
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 17

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.


Experiment series 18

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: 0.1
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: 4.15


Experiment series 19

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: 0.04
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
The affinity was below the detection threshold of the assay.


Experiment series 20

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.


Experiment series 21

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: -0.22
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 22

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

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
The affinity was below the detection threshold of the assay.


Experiment series 23

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 24

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

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


Experiment series 25

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: 0.18
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: 4.1


Experiment series 26

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.


Experiment series 27

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: 0.12
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: 4.15


Experiment series 28

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: -0.02
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
The affinity was below the detection threshold of the assay.


Experiment series 29

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: POM121C
Protein B fragment: 127-141
Protein B site: PxxP_1
Protein B sequence: YLGKPGPPQPAPAPE
Protein B construct: Bio Ado Ado-YLGKPGPPQPAPAPE-CONH2

Average holdup BI: 0.02
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 30

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: POM121C
Protein B fragment: 148-162
Protein B site: PxxP_2
Protein B sequence: RPGRRPPARPAPRST
Protein B construct: Bio Ado Ado-RPGRRPPARPAPRST-CONH2

Average holdup BI: 0.22
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: 4.22


Experiment series 31

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: POM121C
Protein B fragment: 419-433
Protein B site: PxxP_3
Protein B sequence: SSRSQTPERPAKKIR
Protein B construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2

Average holdup BI: 0.15
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: 3.99