Details of experiments between Q04917 and Q96FS4
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: SIPA1
Protein B fragment: 903-912
Protein B site: 14-3-3_pSer908
Protein B modification: (SEP)908
Protein B sequence: LPRTLSLRNS
Protein B construct: LPRTL(pSer)LRNS (crude synthetic peptide)
Average holdup BI: 0.5
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.42
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: 3.89
Experiment series 2
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: SIPA1
Protein B fragment: 832-841
Protein B site: 14-3-3_pSer837
Protein B modification: (SEP)837
Protein B sequence: LHSQNSLSPR
Protein B construct: LHSQN(pSer)LSPR (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.57
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.
Experiment series 3
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: SIPA1
Protein B fragment: 50-59
Protein B site: 14-3-3_pSer55
Protein B modification: (SEP)55
Protein B sequence: LLRSGSDAGE
Protein B construct: LLRSG(pSer)DAGE (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.27
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.3
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.
Experiment series 4
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: SIPA1
Protein B fragment: 554-563
Protein B site: 14-3-3_pSer559
Protein B modification: (SEP)559
Protein B sequence: RFGLPSLGGR
Protein B construct: RFGLP(pSer)LGGR (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.39
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.21
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.