Details of experiments between Q14155 and P30260
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: CDC27
Protein B fragment: 1-824
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.19
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): 5.08
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
The affinity was below the detection threshold of the assay.
Experiment series 2
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: CDC27
Protein B fragment: 396-410
Protein B site: PxxP_1
Protein B sequence: LKMKFPPKIPNRKTK
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 4
Measured pKd: 4.23
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: CDC27
Protein B fragment: 396-410
Protein B site: PxxP_1
Protein B sequence: LKMKFPPKIPNRKTK
Protein B construct: Bio E E Ado Ado-LKMKFPPKIPNRKTK-CONH2
Average holdup BI: 0.16
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: 4.04