Details of experiments between Q8WX93 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: PALLS
Protein A fragment: 1374-1383
Protein A site: PBM
Protein A sequence: NTALVESEDL
Protein A construct: biotin-ado-ado-NTALVESEDL
Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.29
Normalized holdup BI: 0.29
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: 4.38
Experiment series 2
Protein A protein: PALLD
Protein A fragment: 1-1383
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: 0.53
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): 2.59
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
Measured pKd: 5.06
Experiment series 3
Protein A protein: PALLD
Protein A fragment: 1-1383
Protein A construct: endogenous protein (SH-SY5Y)
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: 0.51
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): 3.35
Experiment method: nHU_SH-SY5Y
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
Measured pKd: 5.02
Experiment series 4
Protein A protein: PALLD
Protein A fragment: 1-1383
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: 0.55
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): 6.15
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: 5.09