Details of experiments between O43426 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: SYNJ1
Protein A fragment: 1-1573
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: 0.55
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: SYNJ1
Protein A fragment: 1056-1070
Protein A site: PxxP_1
Protein A sequence: ISEGPVPSLPIRPSR
Protein A construct: Bio Ado Ado-ISEGPVPSLPIRPSR-CONH2

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

Average holdup BI: 0.25
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: SYNJ1
Protein A fragment: 1059-1073
Protein A site: PxxP_2
Protein A sequence: GPVPSLPIRPSRAPS
Protein A construct: Bio Ado Ado-GPVPSLPIRPSRAPS-CONH2

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

Average holdup BI: 0.06
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: SYNJ1
Protein A fragment: 1096-1110
Protein A site: PxxP_3
Protein A sequence: QKDPAQPLEPKRPPP
Protein A construct: Bio Ado Ado-QKDPAQPLEPKRPPP-CONH2

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

Average holdup BI: 0.05
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: SYNJ1
Protein A fragment: 1104-1118
Protein A site: PxxP_4
Protein A sequence: EPKRPPPPRPVAPPT
Protein A construct: Bio Ado Ado-EPKRPPPPRPVAPPT-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.

Experiment series 6

Protein A protein: SYNJ1
Protein A fragment: 1114-1128
Protein A site: PxxP_5
Protein A sequence: VAPPTRPAPPQRPPP
Protein A construct: Bio Ado Ado-VAPPTRPAPPQRPPP-CONH2

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

Average holdup BI: 0.08
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: SYNJ1
Protein A fragment: 1190-1204
Protein A site: PxxP_6
Protein A sequence: GYSTARPTIPPRAGV
Protein A construct: Bio Ado Ado-GYSTARPTIPPRAGV-CONH2

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

Average holdup BI: 0.2
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: SYNJ1
Protein A fragment: 1278-1292
Protein A site: PxxP_7
Protein A sequence: PNLETPPQPPPRSRS
Protein A construct: Bio Ado Ado-PNLETPPQPPPRSRS-CONH2

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

Average holdup BI: 0.55
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: SYNJ1
Protein A fragment: 1371-1385
Protein A site: PxxP_8
Protein A sequence: SCMPTMPPIPARSQS
Protein A construct: Bio Ado Ado-SCMPTMPPIPARSQS-CONH2

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

Average holdup BI: 0.14
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: SYNJ1
Protein A fragment: 1407-1421
Protein A site: PxxP_9
Protein A sequence: SDRTAAPGNPFRAKS
Protein A construct: Bio Ado Ado-SDRTAAPGNPFRAKS-CONH2

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

Average holdup BI: 0.01
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: SYNJ1
Protein A fragment: 1533-1547
Protein A site: PxxP_10
Protein A sequence: LPSRRPPPPPVPLLP
Protein A construct: Bio Ado Ado-LPSRRPPPPPVPLLP-CONH2

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

Average holdup BI: 0.22
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