Details of experiments between Q96L92 and P23634
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: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
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: 5.58
Experiment series 2
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.7
Immobilized partner concentration in holdup experiment (10-6M): 7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.74
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: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.88
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.88
Normalized holdup BI standard deviation: 0.02
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: 3
Measured pKd: 5.7
Experiment series 4
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.88
Normalized holdup BI: 0.88
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.72
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: ATP2B4
Protein B fragment: 1-1241
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.36
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): 4.03
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: 4.75
Experiment series 6
Protein A protein: SNX27
Protein A fragment: 1-541
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SNX27
Protein B protein: ATP2B4
Protein B fragment: 1-1241
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.48
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): 5.8
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: 4.97
Experiment series 7
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: ATP2B4
Protein B fragment: 1-1241
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.3
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): 4.14
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
The affinity was below the detection threshold of the assay.