Details of experiments between the fragments Q6ZMN7~214-318 and O75084~565-574
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PDZRN4
Protein A fragment: 214-318
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FZD7
Protein B fragment: 565-574
Protein B site: PBM
Protein B sequence: SHSSKGETAV
Protein B construct: biotin-ttds-SHSSKGETAV
Measured dissociation constant (10-6M): 105
Standard deviation of dissociation constant (10-6M): 8.49
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.98
Experiment series 2
Protein A protein: PDZRN4
Protein A fragment: 214-318
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FZD7
Protein B fragment: 565-574
Protein B site: PBM
Protein B sequence: SHSSKGETAV
Protein B construct: biotin-ttds-SHSSKGETAV
Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: PDZRN4
Protein A fragment: 214-318
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FZD7
Protein B fragment: 565-574
Protein B site: PBM
Protein B sequence: SHSSKGETAV
Protein B construct: biotin-ttds-SHSSKGETAV
Average holdup BI: 0.29
Normalized holdup BI: 0.29
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4.3