Details of experiments between the fragments P62258~1-255 and Q7KZI7~395-404
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: MARK2
Protein B fragment: 395-404
Protein B site: 14-3-3_pSer400
Protein B modification: (SEP)400
Protein B sequence: VQRSVSANPK
Protein B construct: VQRSV(pSer)ANPK (crude synthetic peptide)
Average holdup BI: 0.47
Holdup BI standard deviation: 0.15
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): 2.36
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: 3.88
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: MARK2
Protein B fragment: 395-404
Protein B site: 14-3-3_pSer400
Protein B modification: (SEP)400
Protein B sequence: VQRSVSANPK
Protein B construct: VQRSV(pSer)ANPK (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 1.48
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): 0.05
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.