Interaction Details


Details of experiments between Q96J02 and Q14155
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: ARHGEF7
Fragment: 178-251
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.92
Immobilized partner concentration (10-6M): 10
P value: 12.95
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-29
Number of measurements: 3
Measured pKd: 6.06
PUBMED ID: 38995680
Experimental Details

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

Experiment series 2

Protein A

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

Protein B

Protein: ARHGEF7
Fragment: 178-251
Site: SH3
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.54
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 pKd: 4.86
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: ARHGEF7
Fragment: 178-251
Site: SH3
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.22
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 pKd: 4.22
PUBMED ID: 38995680

Experiment series 4

Protein A

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

Protein B

Protein: ARHGEF7
Fragment: 178-251
Site: SH3
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.14
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 pKd: 3.97
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: ARHGEF7
Fragment: 178-251
Site: SH3
Construct: his6-MBP-TEVsite-SH3
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