Details of experiments between Q96L92 and Q14155
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: 1-541
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0.09
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): 1.01
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: SNX27
Protein A fragment: 1-541
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ARHGEF7
Protein B fragment: 794-803
Protein B site: PBM
Protein B sequence: NDPAWDETNL
Protein B construct: biotin-ttds-NDPAWDETNL
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
PUBMED ID of publication: 36542723
Number of measurements: 3
Measured pKd: 5.82
Experiment series 3
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF7
Protein B fragment: 794-803
Protein B site: PBM
Protein B sequence: NDPAWDETNL
Protein B construct: biotin-ttds-NDPAWDETNL
Average holdup BI: 0.87
Holdup BI standard deviation: 0
Normalized holdup BI: 0.87
Normalized holdup BI standard deviation: 0
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
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 5.66
Experiment series 4
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: ARHGEF7
Protein B fragment: 1-803
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.8
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): 2.97
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: 5.61
Experiment series 5
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: ARHGEF7
Protein B fragment: 1-803
Protein B construct: endogenous protein (SH-SY5Y)
Average holdup BI: 0.74
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): 2.97
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: 5.46
Experiment series 6
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: ARHGEF7
Protein B fragment: 1-803
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.69
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): 2.4
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: 5.36