Details of experiments between the fragments P31946~1-246 and Q92974~117-126
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: ARHGEF2
Protein B fragment: 117-126
Protein B site: 14-3-3_pSer122
Protein B modification: (SEP)122
Protein B sequence: RERPSSAIYP
Protein B construct: RERPS(pSer)AIYP (crude synthetic peptide)
Average holdup BI: 0.49
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.98
Experiment series 2
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: ARHGEF2
Protein B fragment: 117-126
Protein B site: 14-3-3_pSer122
Protein B modification: (SEP)122
Protein B sequence: RERPSSAIYP
Protein B construct: RERPS(pSer)AIYP (crude synthetic peptide)
Average holdup BI: 0.44
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.64
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.77