Details of experiments between the fragments O15018~326-424 and O15439~1316-1325
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PDZD2
Protein A fragment: 326-424
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 10
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 2
Protein A protein: PDZD2
Protein A fragment: 326-424
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.65
Experiment series 3
Protein A protein: PDZD2
Protein A fragment: 326-424
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.18
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: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 4.11
Experiment series 4
Protein A protein: PDZD2
Protein A fragment: 326-424
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: Biotin-ado-ado-STLTIFETAL
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: 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: 3.59