Details of experiments between Q5TCQ9 and P03146
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: 0.78
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal 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: 2
Measured pKd: 5.37
Experiment series 2
Protein A protein: MAGI3
Protein A fragment: 1-114
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: -0.05
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: MAGI3
Protein A fragment: 572-655
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: MAGI3
Protein A fragment: 716-815
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: -0.12
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: MAGI3
Protein A fragment: 847-941
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: -0.01
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: MAGI3
Protein A fragment: 1014-1103
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: -0
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HBV-Hbc
Protein B fragment: 174-183
Protein B site: PBM
Protein B sequence: RRSQSRESQC
Protein B construct: biotin-(PEG)3-RRRRSQSRESQC
Average holdup BI: 0.4
Normalized holdup BI: 0.42
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: 4.64