Details of experiments between P78352 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: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
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: 0.25
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.45
Experiment series 2
Protein A protein: DLG4
Protein A fragment: 306-403
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: 0.14
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.16
Normalized holdup BI standard deviation: 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
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 4.03
Experiment series 3
Protein A protein: DLG4
Protein A fragment: 62-152
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: 0.04
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
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: DLG4
Protein A fragment: 306-403
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: 0.17
Normalized holdup BI: 0.2
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: 4.15
Experiment series 5
Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
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: 0.3
Normalized holdup BI: 0.38
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: 4.57
Experiment series 6
Protein A protein: DLG4
Protein A fragment: 62-152
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: 0.16
Normalized holdup BI: 0.18
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: 4.11