Skip Navigation


JXB Advance Access originally published online on March 30, 2006
Journal of Experimental Botany 2006 57(7):1515-1522; doi:10.1093/jxb/erj163
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
57/7/1515    most recent
erj163v1
Right arrow E-letters: Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when E-letters are posted
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 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 (7)
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Whitelegge, J. P
Right arrow Articles by Cramer, W. A
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Whitelegge, J. P
Right arrow Articles by Cramer, W. A
Agricola
Right arrow Articles by Whitelegge, J. P
Right arrow Articles by Cramer, W. A
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Sequencing covalent modifications of membrane proteins

Julian P Whitelegge1,*, Arthur Laganowsky2, John Nishio2, Puneet Souda1, Huamin Zhang3 and William A Cramer3

1The Pasarow Mass Spectrometry Laboratory, The Jane and Terry Semel Institute for Neuroscience and Human Behavior, The Brain Research Institute and The Molecular Biology Institute, David Geffen School of Medicine, University of California, 405 Hilgard Avenue, Los Angeles, CA 90095, USA
2College of Natural Sciences, California State University, Chico, California, USA
3Department of Biological Sciences, Purdue University, West Lafayette, Indiana, USA

*To whom correspondence should be addressed. E-mail: jpw{at}chem.ucla.edu

A number of strategies have successfully extended plant proteomics into the bilayer domain. Important benefits can be afforded by including a well-resolved intact protein mass spectrum alongside peptide identification experiments; recent studies of thylakoid membranes have yielded new information on the primary structure and covalent post-translational modification of many of the integral proteins. Intact mass proteomics is advancing through the development of core technologies in separations and mass spectrometry, with the goal of providing comprehensive primary structure coverage that includes transmembrane domains with reliable measurements of protein abundance and turnover. To address the limitations associated with separation technologies such as 2D-gel electrophoresis, alternative systems are being investigated and 2D-liquid chromatography of thylakoid membrane proteins, using both denaturing and non-denaturing first dimensions, has been successful, extending separation space and providing intact protein solutions for electrospray-ionization mass spectrometry and top-down proteomics. High-resolution conventional, and Fourier-transform, mass spectrometry is bringing increasing resolution to tandem mass spectrometry allowing for ‘top-down’ mass spectrometry of intact proteins. Thus the core chromatographic technologies already developed for intact mass proteomics of integral membrane proteins also allow their ‘top-down’ analysis. Thylakoid membrane proteins with one and two transmembrane helices have been analysed demonstrating the ability of collision-activated dissociation (CAD) to sequence through transmembrane domains. When analysed by top-down proteomics, the small subunits of the cytochrome b6f complex from Arabidopsis thylakoids reveal novel post-transcriptional/translational modifications including the presence of glutamic acid at position 2 of PetL instead of the proline residue predicted from the gene sequence.

Key words: 2D-chromatography, bilayer, chloroplast, integral membrane protein, proteome, proteomics, thylakoid, top-down


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
 J Exp BotHome page
A. M. Timperio, G. M. D'Amici, C. Barta, F. Loreto, and L. Zolla
Proteomic, pigment composition, and organization of thylakoid membranes in iron-deficient spinach leaves
J. Exp. Bot., October 10, 2007; (2007) erm219v1.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. Carroll, M. C. Altman, I. M. Fearnley, and J. E. Walker
Identification of membrane proteins by tandem mass spectrometry of protein ions
PNAS, September 4, 2007; 104(36): 14330 - 14335.
[Abstract] [Full Text] [PDF]

Home page
 Brief Funct Genomic ProteomicHome page
K. J. Grant and C. C. Wu
Advances in neuromembrane proteomics: efforts towards a comprehensive analysis of membrane proteins in the brain
Brief Funct Genomic Proteomic, March 12, 2007; (2007) elm001v1.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. M. Taylor, D. Baniulis, J. B. Burritt, J. M. Gripentrog, C. I. Lord, M. H. Riesselman, W. S. Maaty, B. P. Bothner, T. E. Angel, E. A. Dratz, et al.
Analysis of Human Phagocyte Flavocytochrome b558 by Mass Spectrometry
J. Biol. Chem., December 1, 2006; 281(48): 37045 - 37056.
[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.