Details of experiments between the fragments Q13884~107-196 and Q9UHC3~522-531
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.05
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
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.23
Normalized holdup BI: 0.27
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: 4.07
Experiment series 3
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: Biotin-ado-ado-HRTCYLVTQL
Average holdup BI: 0.19
Normalized holdup BI: 0.21
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: 4.19