Details of experiments between Q14160 and Q5VV41
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: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Measured dissociation constant (10-6M): 20.39
Standard deviation of dissociation constant (10-6M): 2.17
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.69
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.34
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: 4.78
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: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.2
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: 4.42
Experiment series 4
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.17
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: 4.33
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: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.05
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
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: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.63
Normalized holdup BI: 0.7
Immobilized partner concentration in holdup experiment (10-6M): 17
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: 5.21
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: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.48
Normalized holdup BI: 0.6
Immobilized partner concentration in holdup experiment (10-6M): 17
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: 5.01
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: ARHGEF16
Protein B fragment: 700-709
Protein B site: PBM
Protein B sequence: MERLRVETDV
Protein B construct: biotin-ttds-MERLRVETDV
Average holdup BI: 0.35
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.37
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 17
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: 4.58