Details of experiments between the fragments P31947~1-248 and Q8WUI4~150-159
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.76
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.2
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: 4.46
Experiment series 2
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.95
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: 4.52
Experiment series 3
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.83
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.82
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: 4.53