Details of experiments between Q14155 and O43426
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1-1573
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.55
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): 5.31
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: 5.09
Experiment series 2
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1056-1070
Protein B site: PxxP_1
Protein B sequence: ISEGPVPSLPIRPSR
Protein B construct: Bio Ado Ado-ISEGPVPSLPIRPSR-CONH2
Average holdup BI: 0.25
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: 4.28
Experiment series 3
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1059-1073
Protein B site: PxxP_2
Protein B sequence: GPVPSLPIRPSRAPS
Protein B construct: Bio Ado Ado-GPVPSLPIRPSRAPS-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: 3.59
Experiment series 4
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1096-1110
Protein B site: PxxP_3
Protein B sequence: QKDPAQPLEPKRPPP
Protein B construct: Bio Ado Ado-QKDPAQPLEPKRPPP-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1104-1118
Protein B site: PxxP_4
Protein B sequence: EPKRPPPPRPVAPPT
Protein B construct: Bio Ado Ado-EPKRPPPPRPVAPPT-CONH2
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.
Experiment series 6
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1114-1128
Protein B site: PxxP_5
Protein B sequence: VAPPTRPAPPQRPPP
Protein B construct: Bio Ado Ado-VAPPTRPAPPQRPPP-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: 2023/01/17
Number of measurements: 1
Measured pKd: 3.7
Experiment series 7
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1190-1204
Protein B site: PxxP_6
Protein B sequence: GYSTARPTIPPRAGV
Protein B construct: Bio Ado Ado-GYSTARPTIPPRAGV-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: 2023/01/17
Number of measurements: 1
Measured pKd: 4.15
Experiment series 8
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1278-1292
Protein B site: PxxP_7
Protein B sequence: PNLETPPQPPPRSRS
Protein B construct: Bio Ado Ado-PNLETPPQPPPRSRS-CONH2
Average holdup BI: 0.55
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: 4.89
Experiment series 9
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1371-1385
Protein B site: PxxP_8
Protein B sequence: SCMPTMPPIPARSQS
Protein B construct: Bio Ado Ado-SCMPTMPPIPARSQS-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: 2023/01/17
Number of measurements: 1
Measured pKd: 3.97
Experiment series 10
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1407-1421
Protein B site: PxxP_9
Protein B sequence: SDRTAAPGNPFRAKS
Protein B construct: Bio Ado Ado-SDRTAAPGNPFRAKS-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 11
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ1
Protein B fragment: 1533-1547
Protein B site: PxxP_10
Protein B sequence: LPSRRPPPPPVPLLP
Protein B construct: Bio Ado Ado-LPSRRPPPPPVPLLP-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
Measured pKd: 4.22