Details of experiments between the fragments Q7Z628~587-596 and Q13424~84-171
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.45
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: 4.39
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 80.25
Standard deviation of dissociation constant (10-6M): 5.31
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.1
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.2
Holdup BI standard deviation: 0.15
Normalized holdup BI: 0.23
Normalized holdup BI standard deviation: 0.17
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: 4.25
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.09
Normalized holdup BI: 0.12
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: 3.9
Experiment series 5
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.37
Normalized holdup BI: 0.41
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: 4.64