Details of experiments between Q86UL8 and Q9P2E8
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: MARCHF4
Protein B fragment: 401-410
Protein B site: PBM
Protein B sequence: RELVMRVTTV
Protein B construct: biotin-ado-ado-RELVMRVTTV
Average holdup BI: 0.6
Normalized holdup BI: 0.67
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: 5.12
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: MARCHF4
Protein B fragment: 401-410
Protein B site: PBM
Protein B sequence: RELVMRVTTV
Protein B construct: biotin-ado-ado-RELVMRVTTV
Average holdup BI: 0.38
Normalized holdup BI: 0.51
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: 4.82
Experiment series 3
Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MARCHF4
Protein B fragment: 401-410
Protein B site: PBM
Protein B sequence: RELVMRVTTV
Protein B construct: biotin-ado-ado-RELVMRVTTV
Average holdup BI: 0.35
Normalized holdup BI: 0.37
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: 4.56
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: MARCHF4
Protein B fragment: 401-410
Protein B site: PBM
Protein B sequence: RELVMRVTTV
Protein B construct: biotin-ado-ado-RELVMRVTTV
Average holdup BI: 0.37
Normalized holdup BI: 0.37
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: 4.55
Experiment series 5
Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MARCHF4
Protein B fragment: 401-410
Protein B site: PBM
Protein B sequence: RELVMRVTTV
Protein B construct: biotin-ado-ado-RELVMRVTTV
Average holdup BI: 0.16
Normalized holdup BI: 0.16
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: 4.05