Details of experiments between the fragments P57105~6-103 and P25054~2834-2843
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: APC
Protein B fragment: 2834-2843
Protein B site: PBM
Protein B sequence: HSGSYLVTSV
Protein B construct: Biotin-Ahx-HSGSYLVTSV
Average holdup BI: 0.42
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.51
Normalized holdup BI standard deviation: 0.02
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: The peptide stock solution had solubility issues. 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: 4.81
Experiment series 2
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: APC
Protein B fragment: 2834-2843
Protein B site: PBM
Protein B sequence: HSGSYLVTSV
Protein B construct: Biotin-Ahx-HSGSYLVTSV
Average holdup BI: 0.58
Normalized holdup BI: 0.61
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.01