Details of experiments between the fragments Q53ET0~65-74 and P31947~1-248
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: CRTC2
Protein A fragment: 65-74
Protein A site: 14-3-3_pSer70
Protein A modification: (SEP)70
Protein A sequence: SHYGGSLPNV
Protein A construct: SHYGG(pSer)LPNV (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.53
Holdup BI standard deviation: 0.17
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): 3.32
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: 3.9
Experiment series 2
Protein A protein: CRTC2
Protein A fragment: 65-74
Protein A site: 14-3-3_pSer70
Protein A modification: (SEP)70
Protein A sequence: SHYGGSLPNV
Protein A construct: SHYGG(pSer)LPNV (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.57
Holdup BI standard deviation: 0.34
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): 1.65
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: 4.06