Details of experiments between the fragments P23634~1232-1241 and Q14160~718-814
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ATP2B4
Protein A fragment: 1232-1241
Protein A site: PBM
Protein A sequence: SSLQSLETSV
Protein A construct: biotin-ttds-SSLQSLETSV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 9.44
Standard deviation of dissociation constant (10-6M): 0.59
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.03
Experiment series 2
Protein A protein: ATP2B4
Protein A fragment: 1232-1241
Protein A site: PBM
Protein A sequence: SSLQSLETSV
Protein A construct: biotin-ttds-SSLQSLETSV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.43
Immobilized partner concentration in holdup experiment (10-6M): 7
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: 5.15
Experiment series 3
Protein A protein: ATP2B4
Protein A fragment: 1232-1241
Protein A site: PBM
Protein A sequence: SSLQSLETSV
Protein A construct: biotin-ttds-SSLQSLETSV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.59
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.62
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
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: 5.03
Experiment series 4
Protein A protein: ATP2B4
Protein A fragment: 1232-1241
Protein A site: PBM
Protein A sequence: SSLQSLETSV
Protein A construct: biotin-ttds-SSLQSLETSV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.66
Normalized holdup BI: 0.66
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.1