Details of experiments between A6NIM6 and Q9H5P4
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SLC15A5
Protein A fragment: 570-579
Protein A site: PBM
Protein A sequence: SSIDLWETAL
Protein A construct: biotin-ttds-SSIDLWETAL
Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.48
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: 5.75
Experiment series 2
Protein A protein: SLC15A5
Protein A fragment: 570-579
Protein A site: PBM
Protein A sequence: SSIDLWETAL
Protein A construct: biotin-ttds-SSIDLWETAL
Protein B protein: PDZD7
Protein B fragment: 82-170
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.19
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: 4.95
Experiment series 3
Protein A protein: SLC15A5
Protein A fragment: 570-579
Protein A site: PBM
Protein A sequence: SSIDLWETAL
Protein A construct: biotin-ttds-SSIDLWETAL
Protein B protein: PDZD7
Protein B fragment: 199-313
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.03
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
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: SLC15A5
Protein A fragment: 570-579
Protein A site: PBM
Protein A sequence: SSIDLWETAL
Protein A construct: biotin-ttds-SSIDLWETAL
Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.59
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.74
Normalized holdup BI standard deviation: 0.01
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: 2
Measured pKd: 5.56
Experiment series 5
Protein A protein: SLC15A5
Protein A fragment: 570-579
Protein A site: PBM
Protein A sequence: SSIDLWETAL
Protein A construct: biotin-ttds-SSIDLWETAL
Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.57
Normalized holdup BI: 0.68
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: 5.41
Experiment series 6
Protein A protein: SLC15A5
Protein A fragment: 570-579
Protein A site: PBM
Protein A sequence: SSIDLWETAL
Protein A construct: Biotin-ado-ado-SSIDLWETAL
Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.65
Normalized holdup BI: 0.69
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: 5.16