Details of experiments between Q14160 and O43182
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: ARHGAP6
Protein B fragment: 965-974
Protein B site: PBM
Protein B sequence: NPDALPETLV
Protein B construct: biotin-ttds-NPDALPETLV
Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
The affinity was below the detection threshold of the assay.
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: ARHGAP6
Protein B fragment: 965-974
Protein B site: PBM
Protein B sequence: NPDALPETLV
Protein B construct: biotin-ttds-NPDALPETLV
Average holdup BI: 0.14
Normalized holdup BI: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 18.9
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: 3.99
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: ARHGAP6
Protein B fragment: 965-974
Protein B site: PBM
Protein B sequence: NPDALPETLV
Protein B construct: biotin-ttds-NPDALPETLV
Average holdup BI: 0.08
Normalized holdup BI: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 18.9
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: 3.8
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: ARHGAP6
Protein B fragment: 965-974
Protein B site: PBM
Protein B sequence: NPDALPETLV
Protein B construct: biotin-ttds-NPDALPETLV
Average holdup BI: -0
Holdup BI standard deviation: 0.05
Normalized holdup BI: -0
Normalized holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 18.9
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.