Details of experiments between the fragments O15439~1316-1325 and Q8TCX5~504-602
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: RHPN1
Protein B fragment: 504-602
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.49
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: 5.09
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: RHPN1
Protein B fragment: 504-602
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.54
Normalized holdup BI: 0.54
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: 4.87
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: RHPN1
Protein B fragment: 504-602
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.64
Normalized holdup BI: 0.64
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: 5.06
Experiment series 4
Protein A protein: ABCC4
Protein A fragment: 1316-1325
Protein A site: PBM
Protein A sequence: STLTIFETAL
Protein A construct: Biotin-ado-ado-STLTIFETAL
Protein B protein: RHPN1
Protein B fragment: 504-602
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.6
Normalized holdup BI: 0.6
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: 4.98