Details of experiments between Q6IQ26 and Q8NI35
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 1530-1615
Protein B site: PDZ_9
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.07
Normalized holdup BI: 0.07
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: 3.63
Experiment series 2
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 359-469
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.06
Normalized holdup BI: 0.06
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: 3.53
Experiment series 3
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 113-229
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.02
Normalized holdup BI: 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
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
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 1673-1763
Protein B site: PDZ_10
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0
Normalized holdup BI: 0
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
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 238-335
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.02
Normalized holdup BI: 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
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
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 547-651
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
Normalized holdup BI: 0.04
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
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 673-780
Protein B site: PDZ_5
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Normalized holdup BI: 0.03
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
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 1058-1167
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
Normalized holdup BI: 0.04
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
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: DENND5A
Protein A fragment: 1278-1287
Protein A site: PBM
Protein A sequence: ETSLVKGIDI
Protein A construct: biotin-ado-ado-ETSLVKGIDI
Protein B protein: Inadl
Protein B fragment: 1425-1530
Protein B site: PDZ_8
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Normalized holdup BI: 0.03
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
The affinity was below the detection threshold of the assay.