Details of experiments between the fragments Q9UHC3~522-531 and Q92743~365-480
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.3
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.62
Experiment series 2
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.31
Normalized holdup BI: 0.44
Immobilized partner concentration in holdup experiment (10-6M): 33
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.39
Experiment series 3
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: Biotin-ado-ado-HRTCYLVTQL
Protein B protein: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.56
Normalized holdup BI: 0.69
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: 5.17