Details of experiments between the fragments Q13884~107-196 and Q14CM0~1313-1322
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.53
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: 5.4
Experiment series 2
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.61
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0.73
Normalized holdup BI standard deviation: 0.08
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: 3
Measured pKd: 5.17
Experiment series 3
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: Biotin-ado-ado-RLPKIKETTV
Average holdup BI: 0.71
Normalized holdup BI: 0.79
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: 5.41