Details of experiments between the fragments P63104~1-245 and Q6WCQ1~1011-1020
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: MPRIP
Protein B fragment: 1011-1020
Protein B site: 14-3-3_pSer1016
Protein B modification: (SEP)1016
Protein B sequence: NIRSKSVIEQ
Protein B construct: NIRSK(pSer)VIEQ (crude synthetic peptide)
Average holdup BI: 0.49
Holdup BI standard deviation: 0.03
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): 7.13
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: 3.86
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: MPRIP
Protein B fragment: 1011-1020
Protein B site: 14-3-3_pSer1016
Protein B modification: (SEP)1016
Protein B sequence: NIRSKSLKEG
Protein B construct: NIRSK(pSer)LKEG (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.69
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): 0.04
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.