Details of experiments between the fragments Q5T5U3~47-159 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: ARHGAP21
Protein A fragment: 47-159
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: 0.21
Normalized holdup BI: 0.21
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: 4.2
Experiment series 2
Protein A protein: ARHGAP21
Protein A fragment: 47-159
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: 0.26
Normalized holdup BI: 0.26
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: 4.31