Details of experiments between the fragments P31946~1-246 and Q15276~402-411
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: RABEP1
Protein B fragment: 402-411
Protein B site: 14-3-3_pSer407
Protein B modification: (SEP)407
Protein B sequence: LRRAQSTDSL
Protein B construct: LRRAQ(pSer)TDSL (crude synthetic peptide)
Average holdup BI: 0.66
Holdup BI standard deviation: 0.02
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): 10.3
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: 4.28
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: RABEP1
Protein B fragment: 402-411
Protein B site: 14-3-3_pSer407
Protein B modification: (SEP)407
Protein B sequence: LRRAQSTDSL
Protein B construct: LRRAQ(pSer)TDSL (crude synthetic peptide)
Average holdup BI: 0.65
Holdup BI standard deviation: 0.02
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): 6.39
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: 4.14