Details of experiments between the fragments Q9NS75~337-346 and Q8N448~334-426
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: CYSLTR2
Protein A fragment: 337-346
Protein A site: PBM
Protein A sequence: SVWLRKETRV
Protein A construct: biotin-ttds-SVWLRKETRV
Protein B protein: LNX2
Protein B fragment: 334-426
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0
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: CYSLTR2
Protein A fragment: 337-346
Protein A site: PBM
Protein A sequence: SVWLRKETRV
Protein A construct: biotin-ttds-SVWLRKETRV
Protein B protein: LNX2
Protein B fragment: 334-426
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.12
Normalized holdup BI: 0.13
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: 3.77
Experiment series 3
Protein A protein: CYSLTR2
Protein A fragment: 337-346
Protein A site: PBM
Protein A sequence: SVWLRKETRV
Protein A construct: Biotin-ado-ado-SVWLRKETRV
Protein B protein: LNX2
Protein B fragment: 334-426
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.18
Normalized holdup BI: 0.18
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: 4.11