Details of experiments between the fragments Q86UL8~1141-1229 and K9N5R3~71-82
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: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: 0.52
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 20
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 4.78
Experiment series 2
Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Measured dissociation constant (10-6M): 45.56
Standard deviation of dissociation constant (10-6M): 9.4
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.34
Experiment series 3
Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: 0.39
Normalized holdup BI: 0.47
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.75