Details of experiments between Q5T2W1 and Q6ZMN7
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0
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
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: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PDZRN4
Protein B fragment: 1027-1036
Protein B site: PBM
Protein B sequence: HNAFLSVTTV
Protein B construct: Biotin-Ahx-HNAFLSVTTV
Average holdup BI: 0.07
Normalized holdup BI: 0.07
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: The peptide stock solution had solubility issues. 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: 3.61
Experiment series 3
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PDZRN4
Protein B fragment: 1027-1036
Protein B site: PBM
Protein B sequence: HNAFLSVTTV
Protein B construct: Biotin-Ahx-HNAFLSVTTV
Average holdup BI: 0.04
Normalized holdup BI: 0.05
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: The peptide stock solution had solubility issues. 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.
Experiment series 4
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PDZRN4
Protein B fragment: 1027-1036
Protein B site: PBM
Protein B sequence: HNAFLSVTTV
Protein B construct: Biotin-Ahx-HNAFLSVTTV
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: The peptide stock solution had solubility issues. 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.