Details of experiments between the fragments Q14CM0~1313-1322 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: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 11.72
Standard deviation of dissociation constant (10-6M): 1.95
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.93
Experiment series 2
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.33
Immobilized partner concentration in holdup experiment (10-6M): 6.6
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: 4.96
Experiment series 3
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.6
Holdup BI standard deviation: 0.12
Normalized holdup BI: 0.63
Normalized holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 21.6
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: 5
Measured pKd: 4.95
Experiment series 4
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: Biotin-ado-ado-RLPKIKETTV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.73
Normalized holdup BI: 0.73
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: 5.25