Multiple conformations of full-length p53 detected with single-molecule fluorescence resonance energy transfer
- Publikationstyp:
- Zeitschriftenaufsatz
- Metadaten:
-
- Autoren
- Fang Huang
- Sridharan Rajagopalan
- Giovanni Settanni
- Richard J Marsh
- Daven A Armoogum
- Nick Nicolaou
- Angus J Bain
- Eitan Lerner
- Elisha Haas
- Liming Ying
- Alan R Fersht
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000272553000039&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1073/pnas.0909644106
- Externe Identifier
- Clarivate Analytics Document Solution ID: 529XP
- PubMed Identifier: 19933326
- ISSN
- 0027-8424
- Ausgabe der Veröffentlichung
- 49
- Zeitschrift
- PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
- Schlüsselwörter
- natively disordered
- domain-domain interaction
- quaternary structure
- FRET
- time-resolved
- Paginierung
- 20758 - 20763
- Datum der Veröffentlichung
- 2009
- Status
- Published
- Titel
- Multiple conformations of full-length p53 detected with single-molecule fluorescence resonance energy transfer
- Sub types
- Article
- Ausgabe der Zeitschrift
- 106
Datenquelle: Web of Science (Lite)
- Andere Metadatenquellen:
-
- Abstract
- <jats:p>The tumor suppressor p53 is a member of the emerging class of proteins that have both folded and intrinsically disordered domains, which are a challenge to structural biology. Its N-terminal domain (NTD) is linked to a folded core domain, which has a disordered link to the folded tetramerization domain, which is followed by a disordered C-terminal domain. The quaternary structure of human p53 has been solved by a combination of NMR spectroscopy, electron microscopy, and small-angle X-ray scattering (SAXS), and the NTD ensemble structure has been solved by NMR and SAXS. The murine p53 is reported to have a different quaternary structure, with the N and C termini interacting. Here, we used single-molecule FRET (SM-FRET) and ensemble FRET to investigate the conformational dynamics of the NTD of p53 in isolation and in the context of tetrameric full-length p53 (flp53). Our results showed that the isolated NTD was extended in solution with a strong preference for residues 66–86 forming a polyproline II conformation. The NTD associated weakly with the DNA binding domain of p53, but not the C termini. We detected multiple conformations in flp53 that were likely to result from the interactions of NTD with the DNA binding domain of each monomeric p53. Overall, the SM-FRET results, in addition to corroborating the previous ensemble findings, enabled the identification of the existence of multiple conformations of p53, which are often averaged and neglected in conventional ensemble techniques. Our study exemplifies the usefulness of SM-FRET in exploring the dynamic landscape of multimeric proteins that contain regions of unstructured domains.</jats:p>
- Autoren
- Fang Huang
- Sridharan Rajagopalan
- Giovanni Settanni
- Richard J Marsh
- Daven A Armoogum
- Nick Nicolaou
- Angus J Bain
- Eitan Lerner
- Elisha Haas
- Liming Ying
- Alan R Fersht
- DOI
- 10.1073/pnas.0909644106
- eISSN
- 1091-6490
- ISSN
- 0027-8424
- Ausgabe der Veröffentlichung
- 49
- Zeitschrift
- Proceedings of the National Academy of Sciences
- Sprache
- en
- Online publication date
- 2009
- Paginierung
- 20758 - 20763
- Datum der Veröffentlichung
- 2009
- Status
- Published
- Herausgeber
- Proceedings of the National Academy of Sciences
- Herausgeber URL
- http://dx.doi.org/10.1073/pnas.0909644106
- Datum der Datenerfassung
- 2022
- Titel
- Multiple conformations of full-length p53 detected with single-molecule fluorescence resonance energy transfer
- Ausgabe der Zeitschrift
- 106
Datenquelle: Crossref
- Abstract
- The tumor suppressor p53 is a member of the emerging class of proteins that have both folded and intrinsically disordered domains, which are a challenge to structural biology. Its N-terminal domain (NTD) is linked to a folded core domain, which has a disordered link to the folded tetramerization domain, which is followed by a disordered C-terminal domain. The quaternary structure of human p53 has been solved by a combination of NMR spectroscopy, electron microscopy, and small-angle X-ray scattering (SAXS), and the NTD ensemble structure has been solved by NMR and SAXS. The murine p53 is reported to have a different quaternary structure, with the N and C termini interacting. Here, we used single-molecule FRET (SM-FRET) and ensemble FRET to investigate the conformational dynamics of the NTD of p53 in isolation and in the context of tetrameric full-length p53 (flp53). Our results showed that the isolated NTD was extended in solution with a strong preference for residues 66-86 forming a polyproline II conformation. The NTD associated weakly with the DNA binding domain of p53, but not the C termini. We detected multiple conformations in flp53 that were likely to result from the interactions of NTD with the DNA binding domain of each monomeric p53. Overall, the SM-FRET results, in addition to corroborating the previous ensemble findings, enabled the identification of the existence of multiple conformations of p53, which are often averaged and neglected in conventional ensemble techniques. Our study exemplifies the usefulness of SM-FRET in exploring the dynamic landscape of multimeric proteins that contain regions of unstructured domains.
- Addresses
- Medical Research Council Centre for Protein Engineering, Hills Road, Cambridge CB2 0QH, United Kingdom.
- Autoren
- Fang Huang
- Sridharan Rajagopalan
- Giovanni Settanni
- Richard J Marsh
- Daven A Armoogum
- Nick Nicolaou
- Angus J Bain
- Eitan Lerner
- Elisha Haas
- Liming Ying
- Alan R Fersht
- DOI
- 10.1073/pnas.0909644106
- eISSN
- 1091-6490
- Externe Identifier
- PubMed Identifier: 19933326
- PubMed Central ID: PMC2791586
- Funding acknowledgements
- Medical Research Council: MC_U105474168
- Biotechnology and Biological Sciences Research Council: JF20607/2
- Engineering and Physical Sciences Research Council: EP/C536134/1
- Open access
- false
- ISSN
- 0027-8424
- Ausgabe der Veröffentlichung
- 49
- Zeitschrift
- Proceedings of the National Academy of Sciences of the United States of America
- Schlüsselwörter
- Animals
- Humans
- Mice
- Amino Acids
- Protein Subunits
- X-Ray Diffraction
- Fluorescence Resonance Energy Transfer
- Diffusion
- Protein Structure, Quaternary
- Protein Structure, Tertiary
- Protein Binding
- Models, Molecular
- Time Factors
- Tumor Suppressor Protein p53
- Mutant Proteins
- Scattering, Small Angle
- Sprache
- eng
- Medium
- Print-Electronic
- Online publication date
- 2009
- Paginierung
- 20758 - 20763
- Datum der Veröffentlichung
- 2009
- Status
- Published
- Datum der Datenerfassung
- 2009
- Titel
- Multiple conformations of full-length p53 detected with single-molecule fluorescence resonance energy transfer.
- Sub types
- Research Support, Non-U.S. Gov't
- research-article
- Journal Article
- Ausgabe der Zeitschrift
- 106
Files
https://www.pnas.org/content/pnas/106/49/20758.full.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19933326/pdf/?tool=EBI http://www.pnas.org/cgi/reprint/106/49/20758.pdf https://europepmc.org/articles/PMC2791586?pdf=render
Datenquelle: Europe PubMed Central
- Abstract
- The tumor suppressor p53 is a member of the emerging class of proteins that have both folded and intrinsically disordered domains, which are a challenge to structural biology. Its N-terminal domain (NTD) is linked to a folded core domain, which has a disordered link to the folded tetramerization domain, which is followed by a disordered C-terminal domain. The quaternary structure of human p53 has been solved by a combination of NMR spectroscopy, electron microscopy, and small-angle X-ray scattering (SAXS), and the NTD ensemble structure has been solved by NMR and SAXS. The murine p53 is reported to have a different quaternary structure, with the N and C termini interacting. Here, we used single-molecule FRET (SM-FRET) and ensemble FRET to investigate the conformational dynamics of the NTD of p53 in isolation and in the context of tetrameric full-length p53 (flp53). Our results showed that the isolated NTD was extended in solution with a strong preference for residues 66-86 forming a polyproline II conformation. The NTD associated weakly with the DNA binding domain of p53, but not the C termini. We detected multiple conformations in flp53 that were likely to result from the interactions of NTD with the DNA binding domain of each monomeric p53. Overall, the SM-FRET results, in addition to corroborating the previous ensemble findings, enabled the identification of the existence of multiple conformations of p53, which are often averaged and neglected in conventional ensemble techniques. Our study exemplifies the usefulness of SM-FRET in exploring the dynamic landscape of multimeric proteins that contain regions of unstructured domains.
- Autoren
- Fang Huang
- Sridharan Rajagopalan
- Giovanni Settanni
- Richard J Marsh
- Daven A Armoogum
- Nick Nicolaou
- Angus J Bain
- Eitan Lerner
- Elisha Haas
- Liming Ying
- Alan R Fersht
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/19933326
- DOI
- 10.1073/pnas.0909644106
- eISSN
- 1091-6490
- Externe Identifier
- PubMed Central ID: PMC2791586
- Funding acknowledgements
- Biotechnology and Biological Sciences Research Council: JF20607/2
- Medical Research Council: MC_U105474168
- Ausgabe der Veröffentlichung
- 49
- Zeitschrift
- Proc Natl Acad Sci U S A
- Schlüsselwörter
- Amino Acids
- Animals
- Diffusion
- Fluorescence Resonance Energy Transfer
- Humans
- Mice
- Models, Molecular
- Mutant Proteins
- Protein Binding
- Protein Structure, Quaternary
- Protein Structure, Tertiary
- Protein Subunits
- Scattering, Small Angle
- Time Factors
- Tumor Suppressor Protein p53
- X-Ray Diffraction
- Sprache
- eng
- Country
- United States
- Paginierung
- 20758 - 20763
- PII
- 0909644106
- Datum der Veröffentlichung
- 2009
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2010
- Titel
- Multiple conformations of full-length p53 detected with single-molecule fluorescence resonance energy transfer.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Ausgabe der Zeitschrift
- 106
Datenquelle: PubMed
- Beziehungen:
- Eigentum von