Details of experiments between the fragments P61981~1-247 and Q7Z6B7~935-944
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: SRGAP1
Protein B fragment: 935-944
Protein B site: 14-3-3_pSer940
Protein B modification: (SEP)940
Protein B sequence: IRRSTSSGQY
Protein B construct: IRRST(pSer)SGQY (crude synthetic peptide)
Average holdup BI: 0.56
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.86
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
Experiment series 2
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: SRGAP1
Protein B fragment: 935-944
Protein B site: 14-3-3_pSer940
Protein B modification: (SEP)940
Protein B sequence: IRRSTSSGQY
Protein B construct: IRRST(pSer)SGQY (crude synthetic peptide)
Average holdup BI: 0.6
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.47
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: 4.05