Details of experiments between Q5VWQ0 and Q12959
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RSBN1
Protein A fragment: 793-802
Protein A site: PBM
Protein A sequence: NLQEHSTTSV
Protein A construct: biotin-ado-ado-NLQEHSTTSV
Protein B protein: DLG1
Protein B fragment: 314-412
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.13
Normalized holdup BI: 0.15
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: 3.99
Experiment series 2
Protein A protein: RSBN1
Protein A fragment: 793-802
Protein A site: PBM
Protein A sequence: NLQEHSTTSV
Protein A construct: biotin-ado-ado-NLQEHSTTSV
Protein B protein: DLG1
Protein B fragment: 457-570
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.06
Normalized holdup BI: 0.09
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: 3.76
Experiment series 3
Protein A protein: RSBN1
Protein A fragment: 793-802
Protein A site: PBM
Protein A sequence: NLQEHSTTSV
Protein A construct: biotin-ado-ado-NLQEHSTTSV
Protein B protein: DLG1
Protein B fragment: 221-314
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.05
Normalized holdup BI: 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: 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: 3.5
Experiment series 4
Protein A protein: RSBN1
Protein A fragment: 1-802
Protein A construct: endogenous protein (Jurkat)
Protein B protein: DLG1
Protein B fragment: 221-314
Protein B site: PDZ_1
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: -0.07
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): 1.14
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.
Experiment series 5
Protein A protein: RSBN1
Protein A fragment: 1-802
Protein A construct: endogenous protein (Jurkat)
Protein B protein: DLG1
Protein B fragment: 314-412
Protein B site: PDZ_2
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: -0.05
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): 0.66
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.