Details of experiments between the fragments P61981~1-247 and Q00537~87-96
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CDK17
Protein B fragment: 87-96
Protein B site: 14-3-3_pSer92
Protein B modification: (SEP)92
Protein B sequence: MPRNGSRLDI
Protein B construct: MPRNG(pSer)RLDI (crude synthetic peptide)
Average holdup BI: 0.23
Holdup BI standard deviation: 0.3
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.69
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 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: CDK17
Protein B fragment: 87-96
Protein B site: 14-3-3_pSer92
Protein B modification: (SEP)92
Protein B sequence: MPRNGSRLDI
Protein B construct: MPRNG(pSer)RLDI (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.47
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): 0.16
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.