Details of experiments between Q14160 and Q6ZTQ3
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: 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): 125.54
Standard deviation of dissociation constant (10-6M): 10.89
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.9
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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: 0.06
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: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
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: 0.03
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 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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: 0.05
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 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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: -0.02
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 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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: 0.25
Normalized holdup BI: 0.28
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: 4.3
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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: 0.12
Normalized holdup BI: 0.15
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: 3.95
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: RASSF6
Protein B fragment: 360-369
Protein B site: PBM
Protein B sequence: KLVIKTETTV
Protein B construct: biotin-ttds-KLVIKTETTV
Average holdup BI: 0.13
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.14
Normalized holdup BI standard deviation: 0.05
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: 4
Measured pKd: 3.9