Details of experiments between the fragments P62258~1-255 and P27228~140-149
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: HPV35E6
Protein B fragment: 140-149
Protein B site: 14-3-3_pThr147
Protein B modification: (TPO)147
Protein B sequence: PTRRETEV--
Protein B construct: SKPTRRE(pThr)EV (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.39
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
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: HPV35E6
Protein B fragment: 140-149
Protein B site: 14-3-3_pThr147
Protein B modification: (TPO)147
Protein B sequence: PTRRETEV--
Protein B construct: SKPTRRE(pThr)EV (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.32
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.31
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
The affinity was below the detection threshold of the assay.