Details of experiments between the fragments P31947~1-248 and Q53ET0~442-451
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 construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: CRTC2
Protein B fragment: 442-451
Protein B site: 14-3-3_pSer447
Protein B modification: (SEP)447
Protein B sequence: ADARRSQQQL
Protein B construct: ADARR(pSer)QQQL (crude synthetic peptide)
Average holdup BI: -0.49
Holdup BI standard deviation: 0.75
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): 0.68
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
The affinity was below the detection threshold of the assay.
Experiment series 2
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: CRTC2
Protein B fragment: 442-451
Protein B site: 14-3-3_pSer447
Protein B modification: (SEP)447
Protein B sequence: ADARRSQQQL
Protein B construct: ADARR(pSer)QQQL (crude synthetic peptide)
Average holdup BI: -0.43
Holdup BI standard deviation: 0.83
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): 0.54
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
The affinity was below the detection threshold of the assay.