Details of experiments between Q5TCQ9 and Q9UHC3
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI3
Protein A fragment: 847-941
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.31
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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: 4.64
Experiment series 2
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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: 4.23
Experiment series 3
Protein A protein: MAGI3
Protein A fragment: 1-114
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: MAGI3
Protein A fragment: 572-655
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: MAGI3
Protein A fragment: 716-815
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: MAGI3
Protein A fragment: 1014-1103
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL
Average holdup BI: 0.35
Normalized holdup BI: 0.39
Immobilized partner concentration in holdup experiment (10-6M): 33
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: The peptide stock solution had solubility issues. 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: 4.3
Experiment series 8
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: Biotin-ado-ado-HRTCYLVTQL
Average holdup BI: 0.42
Normalized holdup BI: 0.45
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. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.7