Details of experiments between Q14160 and O75084
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: 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): 77.95
Standard deviation of dissociation constant (10-6M): 5.2
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.11
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: FZD7
Protein B fragment: 565-574
Protein B site: PBM
Protein B sequence: SHSSKGETAV
Protein B construct: biotin-ttds-SHSSKGETAV
Average holdup BI: -0.01
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: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
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: -0.04
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 4
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
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: 0.01
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 5
Protein A protein: SCRIB
Protein A fragment: 1090-1197
Protein A site: PDZ_4
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
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 6
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
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: 0.48
Normalized holdup BI: 0.54
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: 4.78
Experiment series 7
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
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: 0.34
Normalized holdup BI: 0.43
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: 4.58
Experiment series 8
Protein A protein: SCRIB
Protein A fragment: 718-814
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: 0.2
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.21
Normalized holdup BI standard deviation: 0.1
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: 4
Measured pKd: 4.11