Skip Navigation

Nucleic Acids Research 2005 33(1):182-189; doi:10.1093/nar/gki151
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (3783K) Freely available
Right arrow Screen PDF (684K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (20)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Dorywalska, M.
Right arrow Articles by Puglisi, J. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dorywalska, M.
Right arrow Articles by Puglisi, J. D.
Related Collections
Right arrow Nucleic acid modification
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 12 January 2005

© 2005, the authors Nucleic Acids Research, Vol. 33 No. 1 © Oxford University Press 2005; all rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use permissions, please contact journals.permissions{at}oupjournals.org.

Article

Site-specific labeling of the ribosome for single-molecule spectroscopy

Magdalena Dorywalska1, Scott C. Blanchard1,2, Ruben L. Gonzalez, Jr1, Harold D. Kim2, Steven Chu2,* and Joseph D. Puglisi1,*

1 Department of Structural Biology, Stanford University School of Medicine Stanford, CA 94305-5126, USA 2 Department of Physics and Applied Physics, Stanford University Stanford, CA 94305-4060, USA

*To whom correspondence should be addressed. Tel: +1 650 498 4397; Fax: +1 650 723 8464; Email: puglisi{at}stanford.edu

Received October 11, 2004. Revised December 7, 2004. Accepted December 7, 2004.

Single-molecule fluorescence spectroscopy can reveal mechanistic and kinetic details that may not be observed in static structural and bulk biochemical studies of protein synthesis. One approach requires site-specific and stable attachment of fluorophores to the components of translation machinery. Fluorescent tagging of the ribosome is a prerequisite for the observation of dynamic changes in ribosomal conformation during translation using fluorescence methods. Modifications of the ribosomal particle are difficult due to its complexity and high degree of sequence and structural conservation. We have developed a general method to label specifically the prokaryotic ribosome by hybridization of fluorescent oligonucleotides to mutated ribosomal RNA. Functional, modified ribosomes can be purified as a homogenous population, and fluorescence can be monitored from labeled ribosomal complexes immobilized on a derivatized quartz surface.

Correspondence may also be addressed to Steven Chu. Tel: +1 650 723 3571; Fax: +1 650 723 9173; Email: schu{at}LBL.gov


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
A. M. Blanco, L. Rausell, B. Aguado, M. Perez-Alonso, and R. Artero
A FRET-based assay for characterization of alternative splicing events using peptide nucleic acid fluorescence in situ hybridization
Nucleic Acids Res., September 1, 2009; 37(17): e116 - e116.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
R. A. Marshall, M. Dorywalska, and J. D. Puglisi
Irreversible chemical steps control intersubunit dynamics during translation
PNAS, October 7, 2008; 105(40): 15364 - 15369.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. Uemura, R. Iizuka, T. Ueno, Y. Shimizu, H. Taguchi, T. Ueda, J. D. Puglisi, and T. Funatsu
Single-molecule imaging of full protein synthesis by immobilized ribosomes
Nucleic Acids Res., July 1, 2008; 36(12): e70 - e70.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. K. Smalley and S. K. Silverman
Fluorescence of covalently attached pyrene as a general RNA folding probe
Nucleic Acids Res., January 9, 2006; 34(1): 152 - 166.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.