Details of experiments between Q5T2W1 and P53778
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: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: biotin-ttds-GARVSKETPL
Average holdup BI: 0.79
Normalized holdup BI: 0.84
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
Measured pKd: 5.56
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: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: biotin-ttds-GARVSKETPL
Average holdup BI: 0.08
Normalized holdup BI: 0.08
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
Measured pKd: 3.72
Experiment series 3
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: biotin-ttds-GARVSKETPL
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: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: Biotin-ado-ado-GARVSKETPL
Average holdup BI: 0.81
Normalized holdup BI: 0.85
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: 5.61
Experiment series 5
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: Biotin-ado-ado-GARVSKETPL
Average holdup BI: 0.14
Normalized holdup BI: 0.14
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: 3.97
Experiment series 6
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MAPK12
Protein B fragment: 358-367
Protein B site: PBM
Protein B sequence: GARVSKETPL
Protein B construct: Biotin-ado-ado-GARVSKETPL
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
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.