Details of experiments between the fragments Q8NI35~1058-1167 and A6NIM6~570-579
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: Inadl
Protein A fragment: 1058-1167
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC15A5
Protein B fragment: 570-579
Protein B site: PBM
Protein B sequence: SSIDLWETAL
Protein B construct: biotin-ttds-SSIDLWETAL
Average holdup BI: 0.61
Immobilized partner concentration in holdup experiment (10-6M): 3.5
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: 6.18
Experiment series 2
Protein A protein: Inadl
Protein A fragment: 1058-1167
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC15A5
Protein B fragment: 570-579
Protein B site: PBM
Protein B sequence: SSIDLWETAL
Protein B construct: biotin-ttds-SSIDLWETAL
Average holdup BI: 0.72
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.79
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 11
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: 4
Measured pKd: 5.69
Experiment series 3
Protein A protein: InaDl
Protein A fragment: 1058-1167
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC15A5
Protein B fragment: 570-579
Protein B site: PBM
Protein B sequence: SSIDLWETAL
Protein B construct: biotin-ttds-SSIDLWETAL
Average holdup BI: 0.72
Normalized holdup BI: 0.79
Immobilized partner concentration in holdup experiment (10-6M): 11
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: 5.69
Experiment series 4
Protein A protein: InaDl
Protein A fragment: 1058-1167
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC15A5
Protein B fragment: 570-579
Protein B site: PBM
Protein B sequence: SSIDLWETAL
Protein B construct: Biotin-ado-ado-SSIDLWETAL
Average holdup BI: 0.78
Normalized holdup BI: 0.78
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: 5.37