Details of experiments between the fragments O15085~41-126 and Q9NS75~337-346
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGEF11
Protein A fragment: 41-126
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CYSLTR2
Protein B fragment: 337-346
Protein B site: PBM
Protein B sequence: SVWLRKETRV
Protein B construct: biotin-ttds-SVWLRKETRV
Average holdup BI: -0.15
Immobilized partner concentration in holdup experiment (10-6M): 6.7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: ARHGEF11
Protein A fragment: 41-126
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CYSLTR2
Protein B fragment: 337-346
Protein B site: PBM
Protein B sequence: SVWLRKETRV
Protein B construct: biotin-ttds-SVWLRKETRV
Average holdup BI: 0.34
Normalized holdup BI: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 27
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: 4.46
Experiment series 3
Protein A protein: ARHGEF11
Protein A fragment: 41-126
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CYSLTR2
Protein B fragment: 337-346
Protein B site: PBM
Protein B sequence: SVWLRKETRV
Protein B construct: Biotin-ado-ado-SVWLRKETRV
Average holdup BI: 0.19
Normalized holdup BI: 0.31
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: 4.43