Details of experiments between the fragments Q96BN8~343-352 and Q14160~718-814
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: biotin-ttds-PVRVCEETSL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.76
Normalized holdup BI: 0.8
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: 5.44
Experiment series 2
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: Biotin-ado-ado-PVRVCEETSL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.89
Normalized holdup BI: 0.89
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: 5.74