Details of experiments between Q5SQI0 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: ATAT1
Protein A fragment: 1-421
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: 0.86
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): 7.65
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: 5.8
Experiment series 2
Protein A protein: ATAT1
Protein A fragment: 210-224
Protein A site: PxxP_1
Protein A sequence: AAVDPTPAAPARKLP
Protein A construct: Bio Ado Ado-AAVDPTPAAPARKLP-CONH2
Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.09
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: 3.75
Experiment series 3
Protein A protein: ATAT1
Protein A fragment: 293-307
Protein A site: PxxP_2
Protein A sequence: RSLRLCPPHPTARLL
Protein A construct: Bio Ado Ado-RSLRLCPPHPTARLL-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: 3.55