Interaction Details

Details of experiments between O00499 and Q96Q45
Note that the results of all experiments are listed, regardless of the modification states of the fragments.

Experiment series 1

Protein A

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

Protein B

Protein: TMEM237
Fragment: 1-408
Construct: endogenous protein (Jurkat)

Average holdup BI: 0.91

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

P value: 1.88

Experiment method: nHU_Jurkat

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-06-29

Number of measurements: 3

Measured pK_d: 5.99

PUBMED ID: 38995680

Experimental Details

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

Experiment series 2

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: 0.03

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 3

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

Average holdup BI: 0

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 4

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: -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: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 38995680

Experiment series 5

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

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 6

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

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: 2023-01-17

Number of measurements: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 38995680

Experiment series 7

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

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 8

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: 0

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 9

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

Average holdup BI: 0

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 10

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: 0.03

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

Average holdup BI: 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: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 38995680

Experiment series 12

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: -0.06

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 13

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

Average holdup BI: 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: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 38995680

Experiment series 14

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

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 15

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

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: 2023-01-16

Number of measurements: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 38995680

Experiment series 16

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: 0.03

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 17

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

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: 2023-01-18

Number of measurements: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 38995680

Experiment series 18

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: -0.09

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 19

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

Average holdup BI: 0.01

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 20

Protein A

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

Protein B

Protein: TMEM237
Fragment: 15-29
Site: PxxP_1
Sequence: RPPRALPPVPSQDDI
Construct: Bio Ado Ado-RPPRALPPVPSQDDI-CONH2

Average holdup BI: 0

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 21

Protein A

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

Protein B

Protein: TMEM237
Fragment: 70-84
Site: PxxP_2
Sequence: KELKEHPEAPVQRRQ
Construct: Bio Ado Ado-KELKEHPEAPVQRRQ-CONH2

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: 2023-01-18

Number of measurements: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 38995680