Details of experiments between the fragments Q6P0Q8~1102-1193 and A4U7F2~140-149
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: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Measured dissociation constant (10-6M): 11.01
Standard deviation of dissociation constant (10-6M): 1.24
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.96
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: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.41
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: 4.91
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: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.78
Normalized holdup BI: 0.78
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 5.27