Details of experiments between the fragments Q86UL8~410-522 and P23634~1232-1241
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Measured dissociation constant (10-6M): 43.72
Standard deviation of dissociation constant (10-6M): 3.29
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.36
Experiment series 2
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.09
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
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.31
Normalized holdup BI: 0.34
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: 1
Measured pKd: 4.49
Experiment series 4
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.16
Normalized holdup BI: 0.18
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: 4.09