Details of experiments between the fragments Q5T2W1~2-107 and P53778~358-367
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: biotin-ttds-GARVSKETPL
Average holdup BI: 0.79
Normalized holdup BI: 0.84
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.56
Experiment series 2
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: Biotin-ado-ado-GARVSKETPL
Average holdup BI: 0.81
Normalized holdup BI: 0.85
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.61