Details of experiments between the fragments Q6P0Q8~1102-1193 and Q5VV41~700-709
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Measured dissociation constant (10-6M): 3.24
Standard deviation of dissociation constant (10-6M): 0.24
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.49
Experiment series 2
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.49
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.07
Experiment series 3
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.83
Normalized holdup BI: 0.83
Immobilized partner concentration in holdup experiment (10-6M): 17
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: 5.56