Details of experiments between Q68EM7 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: ARHGAP17
Protein A fragment: 872-881
Protein A site: PBM
Protein A sequence: IDNDTESTAL
Protein A construct: biotin-ttds-IDNDTESTAL
Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.59
Normalized holdup BI: 0.59
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: 1
Measured pKd: 4.97
Experiment series 2
Protein A protein: ARHGAP17
Protein A fragment: 1-881
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: 0.65
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): 4.43
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: 5.27
Experiment series 3
Protein A protein: ARHGAP17
Protein A fragment: 1-881
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: 0.72
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): 5.29
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: 5.41
Experiment series 4
Protein A protein: ARHGAP17
Protein A fragment: 1-881
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: 0.77
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): 4.77
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: 5.52
Experiment series 5
Protein A protein: ARHGAP17
Protein A fragment: 1-881
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: 0.49
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): 5.91
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: 4.99
Experiment series 6
Protein A protein: ARHGAP17
Protein A fragment: 872-881
Protein A site: PBM
Protein A sequence: IDNDTESTAL
Protein A construct: biotin-ttds-IDNDTESTAL
Protein B protein: SNX27
Protein B fragment: 1-541
Protein B construct: endogenous protein (Jurkat)
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: 4.93