Details of experiments between Q8TBC3 and Q96L92
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.76
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.84
Normalized holdup BI standard deviation: 0.05
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: 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: 2
Measured pKd: 5.54
Experiment series 2
Protein A protein: SHKBP1
Protein A fragment: 1-707
Protein A construct: endogenous protein (Jurkat)
Protein B protein: SNX27
Protein B fragment: 1-541
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SNX27
Average holdup BI: -0.15
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): 0.16
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: SHKBP1
Protein A fragment: 1-707
Protein A construct: endogenous protein (Jurkat)
Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: 0.9
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): 5.45
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: 5.97