Details of experiments between the fragments Q12959~221-314 and Q7Z628~587-596
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.8
Immobilized partner concentration in holdup experiment (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.09
Experiment series 2
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.55
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.65
Normalized holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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: 2
Measured pKd: 5.09
Experiment series 3
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.52
Normalized holdup BI: 0.67
Immobilized partner concentration in holdup experiment (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 5.13
Experiment series 4
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: Biotin-ado-ado-SGGKRKETLV
Average holdup BI: 0.64
Normalized holdup BI: 0.76
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: 5.31