Details of experiments between A4D2P6 and O15439
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: GRID2IP
Protein A fragment: 272-370
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: 0.27
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
Measured pKd: 4.61
Experiment series 2
Protein A protein: GRID2IP
Protein A fragment: 1-87
Protein A site: PDZ_1
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: 0.01
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 3
Protein A protein: GRID2IP
Protein A fragment: 272-370
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: 0.25
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.28
Normalized holdup BI standard deviation: 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: 2
Measured pKd: 4.36
Experiment series 4
Protein A protein: GRID2IP
Protein A fragment: 1-87
Protein A site: PDZ_1
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: 0.05
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
Measured pKd: 3.48
Experiment series 5
Protein A protein: GRID2IP
Protein A fragment: 1-87
Protein A site: PDZ_1
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: 0.05
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: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 3.48
Experiment series 6
Protein A protein: GRID2IP
Protein A fragment: 272-370
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: 0.21
Normalized holdup BI: 0.23
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: 4.25
Experiment series 7
Protein A protein: GRID2IP
Protein A fragment: 272-370
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: 0.32
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
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: 4.45
Experiment series 8
Protein A protein: GRID2IP
Protein A fragment: 1-87
Protein A site: PDZ_1
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: -0.04
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. 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.