Details of experiments between the fragments P57105~6-103 and P60484~393-403
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 29
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKV
Average holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 36
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + 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: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKV
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 4
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
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 5
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKV
Average holdup BI: 0.06
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.08
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 19
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: 2
Measured pKd: 3.65
Experiment series 6
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.05
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.06
Normalized holdup BI standard deviation: 0.02
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: 2
Measured pKd: 3.61
Experiment series 7
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: -0
Normalized holdup BI: -0
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
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: -0.02
Normalized holdup BI: -0.02
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
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: SYNJ2BP
Protein A fragment: 6-103
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.03
Normalized holdup BI: 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
Experimental details: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.