Details of experiments between the fragments P57105~6-103 and Q96BN8~343-352
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: OTULIN
Protein B fragment: 343-352
Protein B site: PBM
Protein B sequence: PVRVCEETSL
Protein B construct: biotin-ttds-PVRVCEETSL
Average holdup BI: 0.39
Holdup BI standard deviation: 0.13
Normalized holdup BI: 0.47
Normalized holdup BI standard deviation: 0.16
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: 2
Measured pKd: 4.74
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: OTULIN
Protein B fragment: 343-352
Protein B site: PBM
Protein B sequence: PVRVCEETSL
Protein B construct: Biotin-ado-ado-PVRVCEETSL
Average holdup BI: 0.6
Normalized holdup BI: 0.63
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: 5.03