Details of experiments between Q14160 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: SCRIB
Protein A fragment: 718-814
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
Measured dissociation constant (10-6M): 7.29
Standard deviation of dissociation constant (10-6M): 0.47
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: 5.14
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 718-814
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: 0.59
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: 5.19
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.46
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: 4.94
Experiment series 4
Protein A protein: SCRIB
Protein A fragment: 858-949
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: 0.02
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 5
Protein A protein: SCRIB
Protein A fragment: 1090-1197
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: SCRIB
Protein A fragment: 718-814
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: 0.86
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.91
Normalized holdup BI standard deviation: 0.02
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: 4
Measured pKd: 5.54
Experiment series 7
Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.61
Normalized holdup BI: 0.68
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: 4.85
Experiment series 8
Protein A protein: SCRIB
Protein A fragment: 858-949
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: 0.12
Normalized holdup BI: 0.15
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: 3.73
Experiment series 9
Protein A protein: SCRIB
Protein A fragment: 999-1093
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-ado-ado-HRTCYLVTQL
Average holdup BI: 0.54
Normalized holdup BI: 0.64
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: 5.06
Experiment series 10
Protein A protein: SCRIB
Protein A fragment: 718-814
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-ado-ado-HRTCYLVTQL
Average holdup BI: 0.86
Normalized holdup BI: 0.86
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: 5.62
Experiment series 11
Protein A protein: SCRIB
Protein A fragment: 858-949
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: 0.15
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. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4