Interaction Details

Details of experiments between Q96J02 and O00499
Note that the results of all experiments are listed, regardless of the modification states of the fragments.

Experiment series 1

Protein A

Protein: ITCH
Fragment: 1-903
Construct: endogenous protein (Jurkat)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.92

Immobilized partner concentration (10^-6M): 10

P value: 8.66

Experiment method: nHU_Jurkat

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-06-29

Number of measurements: 3

Measured pK_d: 6.05

PUBMED ID: 38995680

Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))

Experiment series 2

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.77

Immobilized partner concentration (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 pK_d: 5.35

PUBMED ID: 38995680

Experiment series 3

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.71

Immobilized partner concentration (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 pK_d: 5.22

PUBMED ID: 38995680

Experiment series 4

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.25

Holdup BI standard deviation: 0.01

Immobilized partner concentration (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 pK_d: 4.28

PUBMED ID: 38995680

Experiment series 5

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.08

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 6

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.26

Immobilized partner concentration (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 pK_d: 4.32

PUBMED ID: 38995680

Experiment series 7

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.15

Immobilized partner concentration (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 pK_d: 4

PUBMED ID: 38995680

Experiment series 8

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.04

Holdup BI standard deviation: 0.01

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 9

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.06

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 10

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.01

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 11

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.01

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 12

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.03

Holdup BI standard deviation: 0.01

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 13

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.06

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 14

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.56

Immobilized partner concentration (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 pK_d: 5.29

PUBMED ID: 38995680

Experiment series 15

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.5

Immobilized partner concentration (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 pK_d: 5.17

PUBMED ID: 38995680

Experiment series 16

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.1

Holdup BI standard deviation: 0.02

Immobilized partner concentration (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 pK_d: 4.17

PUBMED ID: 38995680

Experiment series 17

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.02

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 18

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.75

Immobilized partner concentration (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 pK_d: 5.3

PUBMED ID: 38995680

Experiment series 19

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.72

Immobilized partner concentration (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 pK_d: 5.21

PUBMED ID: 38995680

Experiment series 20

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.3

Holdup BI standard deviation: 0.02

Immobilized partner concentration (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 pK_d: 4.4

PUBMED ID: 38995680

Experiment series 21

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.01

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 22

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.61

Immobilized partner concentration (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 pK_d: 5.32

PUBMED ID: 38995680

Experiment series 23

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.56

Immobilized partner concentration (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 pK_d: 5.2

PUBMED ID: 38995680

Experiment series 24

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.13

Holdup BI standard deviation: 0.01

Immobilized partner concentration (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 pK_d: 4.19

PUBMED ID: 38995680

Experiment series 25

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.09

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 26

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.79

Immobilized partner concentration (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 pK_d: 5.41

PUBMED ID: 38995680

Experiment series 27

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.73

Immobilized partner concentration (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 pK_d: 5.24

PUBMED ID: 38995680

Experiment series 28

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.24

Holdup BI standard deviation: 0.01

Immobilized partner concentration (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 pK_d: 4.28

PUBMED ID: 38995680

Experiment series 29

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.04

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 30

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.74

Immobilized partner concentration (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 pK_d: 5.28

PUBMED ID: 38995680

Experiment series 31

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.69

Immobilized partner concentration (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 pK_d: 5.15

PUBMED ID: 38995680

Experiment series 32

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.27

Holdup BI standard deviation: 0.02

Immobilized partner concentration (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 pK_d: 4.34

PUBMED ID: 38995680

Experiment series 33

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.05

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 34

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.58

Immobilized partner concentration (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 pK_d: 5.12

PUBMED ID: 38995680

Experiment series 35

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.53

Immobilized partner concentration (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 pK_d: 5.01

PUBMED ID: 38995680

Experiment series 36

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.18

Holdup BI standard deviation: 0

Immobilized partner concentration (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 pK_d: 4.24

PUBMED ID: 38995680

Experiment series 37

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.1

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 38

Protein A

Protein: ITCH
Fragment: 244-258
Site: PxxP_1
Sequence: SNGGFKPSRPPRPSR
Construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.69

Immobilized partner concentration (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 pK_d: 5.17

PUBMED ID: 38995680

Experiment series 39

Protein A

Protein: ITCH
Fragment: 247-261
Site: PxxP_2
Sequence: GFKPSRPPRPSRPPP
Construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.63

Immobilized partner concentration (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 pK_d: 5.04

PUBMED ID: 38995680

Experiment series 40

Protein A

Protein: ITCH
Fragment: 255-269
Site: PxxP_3
Sequence: RPSRPPPPTPRRPAS
Construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.03

Holdup BI standard deviation: 0.01

Immobilized partner concentration (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.

PUBMED ID: 38995680

Experiment series 41

Protein A

Protein: ITCH
Fragment: 353-367
Site: PxxP_4
Sequence: TWDRPEPLPPGWERR
Construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.07

Immobilized partner concentration (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.

PUBMED ID: 38995680