Details of experiments between the fragments P62258~1-255 and Q5TCZ1~1012-1021
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: SH3PXD2A
Protein B fragment: 1012-1021
Protein B site: 14-3-3_pSer1017
Protein B modification: (SEP)1017
Protein B sequence: VRRNSSFSTA
Protein B construct: VRRNS(pSer)FSTA (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.17
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): 0.64
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
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: SH3PXD2A
Protein B fragment: 1012-1021
Protein B site: 14-3-3_pSer1017
Protein B modification: (SEP)1017
Protein B sequence: VRRNSSFSTA
Protein B construct: VRRNS(pSer)FSTA (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.27
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): 0.43
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.