Details of experiments between the fragments A6NIM6~570-579 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: SLC15A5
Protein A fragment: 570-579
Protein A site: PBM
Protein A sequence: SSIDLWETAL
Protein A construct: biotin-ttds-SSIDLWETAL
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): 95.62
Standard deviation of dissociation constant (10-6M): 15.61
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.02
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.04
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 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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.17
Holdup BI standard deviation: 0.18
Normalized holdup BI: 0.18
Normalized holdup BI standard deviation: 0.2
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: 4.33
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.04
Normalized holdup BI: -0.04
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
The affinity was below the detection threshold of the assay.
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.01
Normalized holdup BI: 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
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
The affinity was below the detection threshold of the assay.
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.01
Normalized holdup BI: -0.01
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
The affinity was below the detection threshold of the assay.