Details of experiments between the fragments Q6ZMN7~214-318 and Q6ZTQ3~360-369
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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Measured dissociation constant (10-6M): 128.91
Standard deviation of dissociation constant (10-6M): 7.84
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.89
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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: -0.1
Immobilized partner concentration in holdup experiment (10-6M): 9
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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: 0.22
Normalized holdup BI: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 20.7
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: 4.15