Details of experiments between O15056 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: SYNJ2
Protein A fragment: 1-1496
Protein A construct: endogenous protein (Jurkat)

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.77
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): 4.44
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: 5.52

Experiment series 2

Protein A protein: SYNJ2
Protein A fragment: 1098-1112
Protein A site: PxxP_1
Protein A sequence: VPNRPRPPQPPQRPP
Protein A construct: Bio Ado Ado-VPNRPRPPQPPQRPP-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.1
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: 3.78

Experiment series 3

Protein A protein: SYNJ2
Protein A fragment: 1099-1113
Protein A site: PxxP_2
Protein A sequence: PNRPRPPQPPQRPPP
Protein A construct: Bio Ado Ado-PNRPRPPQPPQRPPP-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

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
Measured pKd: 3.63

Experiment series 4

Protein A protein: SYNJ2
Protein A fragment: 1140-1154
Protein A site: PxxP_3
Protein A sequence: QTARLLPGAPQQPPK
Protein A construct: Bio Ado Ado-QTARLLPGAPQQPPK-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

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 5

Protein A protein: SYNJ2
Protein A fragment: 1143-1157
Protein A site: PxxP_4
Protein A sequence: RLLPGAPQQPPKART
Protein A construct: Bio Ado Ado-RLLPGAPQQPPKART-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.04
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 6

Protein A protein: SYNJ2
Protein A fragment: 1219-1233
Protein A site: PxxP_5
Protein A sequence: ETPQAPPLLPRRPPP
Protein A construct: Bio Ado Ado-ETPQAPPLLPRRPPP-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.59
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: 4.97

Experiment series 7

Protein A protein: SYNJ2
Protein A fragment: 1227-1241
Protein A site: PxxP_6
Protein A sequence: LPRRPPPRVPAIKKP
Protein A construct: Bio Ado Ado-LPRRPPPRVPAIKKP-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.28
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: 4.36

Experiment series 8

Protein A protein: SYNJ2
Protein A fragment: 1324-1338
Protein A site: PxxP_7
Protein A sequence: EAPPLVPKVPPRRKK
Protein A construct: Bio Ado Ado-EAPPLVPKVPPRRKK-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.58
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: 4.94

Experiment series 9

Protein A protein: SYNJ2
Protein A fragment: 1442-1456
Protein A site: PxxP_8
Protein A sequence: RSPKRDPIDPVSAGA
Protein A construct: Bio Ado Ado-RSPKRDPIDPVSAGA-CONH2

Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

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