Details of experiments between the fragments Q9NS75~337-346 and Q6ZMN7~214-318
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 17.41
Standard deviation of dissociation constant (10-6M): 0.73
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.76
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.2
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
Measured pKd: 4.64
Experiment series 3
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.59
Normalized holdup BI: 0.59
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: 4.76
Experiment series 4
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.47
Normalized holdup BI: 0.52
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: 4.83