Details of experiments between the fragments Q7Z628~587-596 and Q86UL8~410-522
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: NET1
Protein A fragment: 587-596
Protein A site: PBM
Protein A sequence: SGGKRKETLV
Protein A construct: biotin-ttds-SGGKRKETLV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.95
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: 5.77
Experiment series 2
Protein A protein: NET1
Protein A fragment: 587-596
Protein A site: PBM
Protein A sequence: SGGKRKETLV
Protein A construct: biotin-ttds-SGGKRKETLV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0.82
Standard deviation of dissociation constant (10-6M): 0.19
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 6.09
Experiment series 3
Protein A protein: NET1
Protein A fragment: 587-596
Protein A site: PBM
Protein A sequence: SGGKRKETLV
Protein A construct: biotin-ttds-SGGKRKETLV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.83
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.91
Normalized holdup BI standard deviation: 0.03
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: 5.87
Experiment series 4
Protein A protein: NET1
Protein A fragment: 587-596
Protein A site: PBM
Protein A sequence: SGGKRKETLV
Protein A construct: biotin-ttds-SGGKRKETLV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.72
Normalized holdup BI: 0.86
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: 5.64
Experiment series 5
Protein A protein: NET1
Protein A fragment: 587-596
Protein A site: PBM
Protein A sequence: SGGKRKETLV
Protein A construct: biotin-ttds-SGGKRKETLV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.79
Normalized holdup BI: 0.87
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: 5.66
Experiment series 6
Protein A protein: NET1
Protein A fragment: 587-596
Protein A site: PBM
Protein A sequence: SGGKRKETLV
Protein A construct: Biotin-ado-ado-SGGKRKETLV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.83
Normalized holdup BI: 0.92
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: 5.9