Details of experiments between Q9UHC3 and P78352
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 179.08
Standard deviation of dissociation constant (10-6M): 45.96
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
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: 3
Measured pKd: 3.75
Experiment series 2
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
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 3
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
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: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.06
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: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.18
Normalized holdup BI: 0.23
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: 3.97
Experiment series 6
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.14
Normalized holdup BI: 0.16
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: 3.78
Experiment series 7
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: biotin-ttds-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.1
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.11
Normalized holdup BI standard deviation: 0.05
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: 2
Measured pKd: 3.6
Experiment series 8
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: Biotin-ado-ado-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
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. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.77
Experiment series 9
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: Biotin-ado-ado-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.17
Normalized holdup BI: 0.21
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: 4.2
Experiment series 10
Protein A protein: ASIC3
Protein A fragment: 522-531
Protein A site: PBM
Protein A sequence: HRTCYLVTQL
Protein A construct: Biotin-ado-ado-HRTCYLVTQL
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
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. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.76