Details of experiments between P63104 and Q96Q42
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: ALS2
Protein B fragment: 461-470
Protein B site: 14-3-3_pSer466
Protein B modification: (SEP)466
Protein B sequence: GKKSSSLVDI
Protein B construct: GKKSS(pSer)LVDI (crude synthetic peptide)
Average holdup BI: 0.4
Holdup BI standard deviation: 0.02
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): 8.23
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
Measured pKd: 3.7
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: ALS2
Protein B fragment: 525-534
Protein B site: 14-3-3_pThr530
Protein B modification: (TPO)530
Protein B sequence: RTEVWTWGKG
Protein B construct: RTEVW(pThr)WGKG (crude synthetic peptide)
Average holdup BI: 0.78
Holdup BI standard deviation: 0.04
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): 5.75
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: 4.42
Experiment series 3
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: ALS2
Protein B fragment: 478-487
Protein B site: 14-3-3_pSer483
Protein B modification: (SEP)483
Protein B sequence: GSRRLSLPGL
Protein B construct: GSRRL(pSer)LPGL (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.07
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): 3.81
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: 4.12