Details of experiments between Q96L92 and P43007
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC1A4
Protein B fragment: 523-532
Protein B site: PBM
Protein B sequence: PELESKESVL
Protein B construct: biotin-ado-ado-PELESKESVL
Average holdup BI: 0.8
Normalized holdup BI: 0.8
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.43
Experiment series 2
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: SLC1A4
Protein B fragment: 1-532
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.49
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.4
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.27.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 6))
PUBMED ID of publication: 36542723
Number of measurements: 6
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: SLC1A4
Protein B fragment: 1-532
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.76
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.22
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
Measured pKd: 5.5