Details of experiments between Q5T2W1 and Q9P202
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: NHERF3
Protein A fragment: 510-519
Protein A site: PBM
Protein A sequence: SSSNSEDTEM
Protein A construct: Biotin-Ahx-SSSNSEDTEM
Protein B protein: DFNB31
Protein B fragment: 136-226
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0
Normalized holdup BI: 0
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: NHERF3
Protein A fragment: 510-519
Protein A site: PBM
Protein A sequence: SSSNSEDTEM
Protein A construct: Biotin-Ahx-SSSNSEDTEM
Protein B protein: DFNB31
Protein B fragment: 263-379
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
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: NHERF3
Protein A fragment: 510-519
Protein A site: PBM
Protein A sequence: SSSNSEDTEM
Protein A construct: Biotin-Ahx-SSSNSEDTEM
Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
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
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: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 565-578
Protein B site: Internal_PBM
Protein B construct: Biotin-GSGSGS-VSVDDVRSTSQGLSSY-CONH2
Average holdup BI: -0.06
Normalized holdup BI: -0.06
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
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: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 565-578
Protein B site: Internal_PBM
Protein B construct: Biotin-GSGSGS-VSVDDVRSTSQGLSSY-CONH2
Average holdup BI: -0.1
Normalized holdup BI: -0.1
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 565-578
Protein B site: Internal_PBM
Protein B construct: Biotin-GSGSGS-VSVDDVRSTSQGLSSY-CONH2
Average holdup BI: 0
Normalized holdup BI: 0
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 898-907
Protein B site: PBM
Protein B sequence: FLVTEFNVML
Protein B construct: biotin-ado-ado-FLVTEFNVML
Average holdup BI: 0.52
Normalized holdup BI: 0.54
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.88
Experiment series 8
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 898-907
Protein B site: PBM
Protein B sequence: FLVTEFNVML
Protein B construct: biotin-ado-ado-FLVTEFNVML
Average holdup BI: 0.37
Normalized holdup BI: 0.37
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.55
Experiment series 9
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 898-907
Protein B site: PBM
Protein B sequence: FLVTEFNVML
Protein B construct: biotin-ado-ado-FLVTEFNVML
Average holdup BI: -0.04
Normalized holdup BI: -0.04
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.