Details of experiments between A6NIM6 and Q86UT5
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: NHERF4
Protein B fragment: 106-208
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.1
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: 4.56
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: NHERF4
Protein B fragment: 220-307
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: NHERF4
Protein B fragment: 325-412
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.09
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: NHERF4
Protein B fragment: 464-550
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
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 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: NHERF4
Protein B fragment: 106-208
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.33
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.33
Normalized holdup BI standard deviation: 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
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 4.71
Experiment series 6
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: NHERF4
Protein B fragment: 464-550
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.2
Normalized holdup BI: 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: 1
Measured pKd: 4.4
Experiment series 7
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: NHERF4
Protein B fragment: 220-307
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0
Normalized holdup BI: -0
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: 1
The affinity was below the detection threshold of the assay.
Experiment series 8
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: NHERF4
Protein B fragment: 106-208
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.36
Normalized holdup BI: 0.36
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: 4.76
Experiment series 9
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: NHERF4
Protein B fragment: 220-307
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.02
Normalized holdup BI: -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
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 10
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: NHERF4
Protein B fragment: 464-550
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.17
Normalized holdup BI: 0.17
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: 4.29
Experiment series 11
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: NHERF4
Protein B fragment: 220-307
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.02
Normalized holdup BI: 0.02
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.
Experiment series 12
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: NHERF4
Protein B fragment: 464-550
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.32
Normalized holdup BI: 0.32
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: 4.45
Experiment series 13
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: NHERF4
Protein B fragment: 106-208
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.41
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: 4.62