Details of experiments between the fragments Q6P0Q8~1102-1193 and P03146~174-183
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: 0.21
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 4.2
Experiment series 2
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: 0.24
Normalized holdup BI: 0.24
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.26