Details of experiments between Q6IQ26 and Q86UL8
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: MAGI2
Protein B fragment: 772-865
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.06
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: 3.66
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
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: 3.48
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: MAGI2
Protein B fragment: 600-682
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
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
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: MAGI2
Protein B fragment: 915-1017
Protein B site: PDZ_5
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 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: MAGI2
Protein B fragment: 1141-1229
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.