Protein Science
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
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 Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by WHITELEGGE, J. P.
Right arrow Articles by FAULL, K. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by WHITELEGGE, J. P.
Right arrow Articles by FAULL, K. F.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Protein Science, Vol 7, Issue 6 1423-1430, Copyright © 1998 by Cold Spring Harbor Laboratory Press

ARTICLE

Electrospray-ionization mass spectrometry of intact intrinsic membrane proteins

J. P. WHITELEGGE, C. B. GUNDERSEN and K. F. FAULL
Center for Molecular and Medical Sciences Mass Spectrometry, University of California, Los Angeles, California 90095 Department of Chemistry & Biochemistry, University of California, Los Angeles, California 90095

Membrane proteins drive and mediate many essential cellular processes making them a vital section of the proteome. However, the amphipathic nature of these molecules ensures their detailed structural analysis remains challenging. A versatile procedure for effective electrospray-ionization mass spectrometry (ESI-MS) of intact intrinsic membrane proteins purified using reverse-phase chromatography in aqueous formic acid/isopropanol is presented. The spectra of four examples, bacteriorhodopsin and its apoprotein from Halobacterium and the D1 and D2 reaction-center subunits from spinach thylakoids, achieve mass measurements that are within 0.01% of calculated theoretical values. All of the spectra reveal lesser quantities of other molecular species that can usually be equated with covalently modified subpopulations of these proteins. Our analysis of bovine rhodopsin, the first ESI-MS study of a G-protein coupled receptor, yielded a complex spectrum indicative of extensive molecular heterogeneity. The range of masses measured for the native molecule agrees well with the range calculated based upon variable glycosylation and reveals further heterogeneity arising from other covalent modifications. The technique described represents the most precise way to catalogue membrane proteins and their post-translational modifications. Resolution of the components of protein complexes provides insights into native protein/protein interactions. The apparent retention of structure by bacteriorhodopsin during the analysis raises the potential of obtaining tertiary structure information using more developed ESI-MS experiments.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
B. F. Shaw, H. L. Lelie, A. Durazo, A. M. Nersissian, G. Xu, P. K. Chan, E. B. Gralla, A. Tiwari, L. J. Hayward, D. R. Borchelt, et al.
Detergent-insoluble Aggregates Associated with Amyotrophic Lateral Sclerosis in Transgenic Mice Contain Primarily Full-length, Unmodified Superoxide Dismutase-1
J. Biol. Chem., March 28, 2008; 283(13): 8340 - 8350.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
L. Banci, I. Bertini, A. Durazo, S. Girotto, E. B. Gralla, M. Martinelli, J. S. Valentine, M. Vieru, and J. P. Whitelegge
Metal-free superoxide dismutase forms soluble oligomers under physiological conditions: A possible general mechanism for familial ALS
PNAS, July 3, 2007; 104(27): 11263 - 11267.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. R. Porras-Yakushi, J. P. Whitelegge, and S. Clarke
Yeast Ribosomal/Cytochrome c SET Domain Methyltransferase Subfamily: IDENTIFICATION OF Rpl23ab METHYLATION SITES AND RECOGNITION MOTIFS
J. Biol. Chem., April 27, 2007; 282(17): 12368 - 12376.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. R. Porras-Yakushi, J. P. Whitelegge, and S. Clarke
A Novel SET Domain Methyltransferase in Yeast: Rkm2-DEPENDENT TRIMETHYLATION OF RIBOSOMAL PROTEIN L12ab AT LYSINE 10
J. Biol. Chem., November 24, 2006; 281(47): 35835 - 35845.
[Abstract] [Full Text] [PDF]

Home page
 Plant CellHome page
M. M. Nowaczyk, R. Hebeler, E. Schlodder, H. E. Meyer, B. Warscheid, and M. Rogner
Psb27, a Cyanobacterial Lipoprotein, Is Involved in the Repair Cycle of Photosystem II
PLANT CELL, November 1, 2006; 18(11): 3121 - 3131.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. Carroll, I. M. Fearnley, and J. E. Walker
Definition of the mitochondrial proteome by measurement of molecular masses of membrane proteins
PNAS, October 31, 2006; 103(44): 16170 - 16175.
[Abstract] [Full Text] [PDF]

Home page
 J Exp BotHome page
J. P Whitelegge, A. Laganowsky, J. Nishio, P. Souda, H. Zhang, and W. A Cramer
Sequencing covalent modifications of membrane proteins
J. Exp. Bot., April 1, 2006; 57(7): 1515 - 1522.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. R. Porras-Yakushi, J. P. Whitelegge, T. B. Miranda, and S. Clarke
A Novel SET Domain Methyltransferase Modifies Ribosomal Protein Rpl23ab in Yeast
J. Biol. Chem., October 14, 2005; 280(41): 34590 - 34598.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
M. Lu, L. Zhang, R. S. Maul, M. R. Sartippour, A. Norris, J. Whitelegge, J. Y. Rao, and M. N. Brooks
The Novel Gene EG-1 Stimulates Cellular Proliferation
Cancer Res., July 15, 2005; 65(14): 6159 - 6166.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Weinglass, J. P. Whitelegge, K. F. Faull, and H. R. Kaback
Monitoring Conformational Rearrangements in the Substrate-binding Site of a Membrane Transport Protein by Mass Spectrometry
J. Biol. Chem., October 1, 2004; 279(40): 41858 - 41865.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Kerner, A. M. Distler, P. Minkler, W. Parland, S. M. Peterman, and C. L. Hoppel
Phosphorylation of Rat Liver Mitochondrial Carnitine Palmitoyltransferase-I: EFFECT ON THE KINETIC PROPERTIES OF THE ENZYME
J. Biol. Chem., September 24, 2004; 279(39): 41104 - 41113.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Lengqvist, R. Svensson, E. Evergren, R. Morgenstern, and W. J. Griffiths
Observation of an Intact Noncovalent Homotrimer of Detergent-solubilized Rat Microsomal Glutathione Transferase-1 by Electrospray Mass Spectrometry
J. Biol. Chem., April 2, 2004; 279(14): 13311 - 13316.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
S. M. Gomez, K. Y. Bil', R. Aguilera, J. N. Nishio, K. F. Faull, and J. P. Whitelegge
Transit Peptide Cleavage Sites of Integral Thylakoid Membrane Proteins
Mol. Cell. Proteomics, October 1, 2003; 2(10): 1068 - 1085.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
M. Ferro, D. Salvi, S. Brugiere, S. Miras, S. Kowalski, M. Louwagie, J. Garin, J. Joyard, and N. Rolland
Proteomics of the chloroplast envelope membranes from Arabidopsis thaliana.
Mol. Cell. Proteomics, May 1, 2003; 2(5): 325 - 345.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
L. Zolla, A.-M. Timperio, W. Walcher, and C. G. Huber
Proteomics of Light-Harvesting Proteins in Different Plant Species. Analysis and Comparison by Liquid Chromatography-Electrospray Ionization Mass Spectrometry. Photosystem II
Plant Physiology, January 1, 2003; 131(1): 198 - 214.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
L. Zolla, S. Rinalducci, A. M. Timperio, and C. G. Huber
Proteomics of Light-Harvesting Proteins in Different Plant Species. Analysis and Comparison by Liquid Chromatography-Electrospray Ionization Mass Spectrometry. Photosystem I
Plant Physiology, December 1, 2002; 130(4): 1938 - 1950.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
J. P. Whitelegge, H. Zhang, R. Aguilera, R. M. Taylor, and W. A. Cramer
Full Subunit Coverage Liquid Chromatography Electrospray Ionization Mass Spectrometry (LCMS+) of an Oligomeric Membrane Protein: Cytochrome b6f Complex From Spinach and the Cyanobacterium Mastigocladus Laminosus
Mol. Cell. Proteomics, October 1, 2002; 1(10): 816 - 827.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. P. Whitelegge
Plant proteomics: BLASTing out of a MudPIT
PNAS, September 3, 2002; 99(18): 11564 - 11566.
[Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
S. M. Gomez, J. N. Nishio, K. F. Faull, and J. P. Whitelegge
The Chloroplast Grana Proteome Defined by Intact Mass Measurements from Liquid Chromatography Mass Spectrometry
Mol. Cell. Proteomics, January 1, 2002; 1(1): 46 - 59.
[Abstract] [Full Text] [PDF]

Home page
 Plant CellHome page
J.-B. Peltier, G. Friso, D. E. Kalume, P. Roepstorff, F. Nilsson, I. Adamska, and K. J. van Wijk
Proteomics of the Chloroplast: Systematic Identification and Targeting Analysis of Lumenal and Peripheral Thylakoid Proteins
PLANT CELL, March 1, 2000; 12(3): 319 - 342.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
D. C. Rees, G. Chang, and R. H. Spencer
Crystallographic Analyses of Ion Channels: Lessons and Challenges
J. Biol. Chem., January 14, 2000; 275(2): 713 - 716.
[Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. P. Whitelegge, J. le Coutre, J. C. Lee, C. K. Engel, G. G. Prive, K. F. Faull, and H. R. Kaback
Toward the bilayer proteome, electrospray ionization-mass spectrometry of large, intact transmembrane proteins
PNAS, September 14, 1999; 96(19): 10695 - 10698.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
D. R. Blanco, J. P. Whitelegge, J. N. Miller, and M. A. Lovett
Demonstration by Mass Spectrometry that Purified Native Treponema pallidum Rare Outer Membrane Protein 1 (Tromp1) Has a Cleaved Signal Peptide
J. Bacteriol., August 15, 1999; 181(16): 5094 - 5098.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
L. Montesano-Roditis, D. G. Glitz, R. R. Traut, and P. L. Stewart
Cryo-electron Microscopic Localization of Protein L7/L12 within the Escherichia coli 70 S Ribosome by Difference Mapping and Nanogold Labeling
J. Biol. Chem., April 20, 2001; 276(17): 14117 - 14123.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. Zhang, J. P. Whitelegge, and W. A. Cramer
Ferredoxin:NADP+ Oxidoreductase Is a Subunit of the Chloroplast Cytochrome b6f Complex
J. Biol. Chem., October 5, 2001; 276(41): 38159 - 38165.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 1998 by The Protein Society.