Details of experiments between the fragments Q14160~718-814 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: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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): 175.84
Standard deviation of dissociation constant (10-6M): 13.44
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.75
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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: 0.24
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: 4.55
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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: 0.03
Holdup BI standard deviation: 0.08
Normalized holdup BI: 0.03
Normalized holdup BI standard deviation: 0.08
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: 4
The affinity was below the detection threshold of the assay.