Details of experiments between Q86UL8 and P34998
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV
Average holdup BI: 0.56
Normalized holdup BI: 0.63
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: 5.03
Experiment series 2
Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV
Average holdup BI: 0.23
Normalized holdup BI: 0.31
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: 4.42
Experiment series 3
Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV
Average holdup BI: 0.29
Normalized holdup BI: 0.29
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: 4.38
Experiment series 4
Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV
Average holdup BI: 0.11
Normalized holdup BI: 0.11
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: 3.83
Experiment series 5
Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV
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: 3.61