Details of experiments between the fragments O15439~1316-1325 and Q14160~718-814
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ABCC4
Protein A fragment: 1316-1325
Protein A site: PBM
Protein A sequence: STLTIFETAL
Protein A construct: biotin-ttds-STLTIFETAL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.64
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: 5.38
Experiment series 2
Protein A protein: ABCC4
Protein A fragment: 1316-1325
Protein A site: PBM
Protein A sequence: STLTIFETAL
Protein A construct: biotin-ttds-STLTIFETAL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.9
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.94
Normalized holdup BI standard deviation: 0.06
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: 5
Measured pKd: 6.07
Experiment series 3
Protein A protein: ABCC4
Protein A fragment: 1316-1325
Protein A site: PBM
Protein A sequence: STLTIFETAL
Protein A construct: biotin-ttds-STLTIFETAL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.75
Normalized holdup BI: 0.94
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: 6.06
Experiment series 4
Protein A protein: ABCC4
Protein A fragment: 1316-1325
Protein A site: PBM
Protein A sequence: STLTIFETAL
Protein A construct: biotin-ttds-STLTIFETAL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.83
Normalized holdup BI: 0.92
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: 5.88
Experiment series 5
Protein A protein: ABCC4
Protein A fragment: 1316-1325
Protein A site: PBM
Protein A sequence: STLTIFETAL
Protein A construct: Biotin-ado-ado-STLTIFETAL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.89
Normalized holdup BI: 0.89
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: 5.73