Details of experiments between P63104 and Q5VT97
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: SYDE2
Protein B fragment: 1188-1197
Protein B site: 14-3-3_pSer1193
Protein B modification: (SEP)1193
Protein B sequence: NKLNMSF---
Protein B construct: LNKNKLNM(pSer)F (crude synthetic peptide)
Average holdup BI: 0.93
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.64
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: 5.02
Experiment series 2
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: SYDE2
Protein B fragment: 617-626
Protein B site: 14-3-3_pSer622
Protein B modification: (SEP)622
Protein B sequence: KGGYLSDGDS
Protein B construct: KGGYL(pSer)DGDS (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.21
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.23
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.