Details of experiments between the fragments Q9H8Y8~1-208 and O14924~1438-1447
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: GORASP2
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RGS12
Protein B fragment: 1438-1447
Protein B site: PBM
Protein B sequence: KTSAHHATFV
Protein B construct: Biotin-Ahx-KTSAHHATFV
Average holdup BI: 0.12
Normalized holdup BI: 0.18
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: 4.12
Experiment series 2
Protein A protein: GORASP2
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RGS12
Protein B fragment: 1438-1447
Protein B site: PBM
Protein B sequence: KTSAHHATFV
Protein B construct: Biotin-ado-ado-KTSAHHATFV
Average holdup BI: 0.22
Normalized holdup BI: 0.22
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: 4.21