Skip Navigation



Glycobiology Advance Access published online on November 1, 2002
Glycobiology, doi:10.1093/glycob/cwg008
This Article
Right arrow Advance Access manuscript (PDF) Freely available
Right arrow All Versions of this Article:
13/2/51    most recent
cwg008v1
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 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 arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Zhou, G.-P.
Right arrow Articles by Troy II, F. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zhou, G.-P.
Right arrow Articles by Troy II, F. A.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Submitted on April 16, 2002
Revised on August 27, 2002
Accepted on September 6, 2002

© 2002 Oxford University Press

ORIGINAL ARTICLES

Characterization by NMR and molecular modeling of the binding of Polyisoprenols (PI) and Polyisoprenyl Recognition Sequence (PIRS) peptides: Three-dimensional structure of the complexes reveals sites of specific interactions

Guo-Ping Zhou 1 Frederic A. Troy II 1*

1 From the Department of Biological Chemistry, University of California School of Medicine, Davis, California 95616

* To whom correspondence should be addressed. E-mail: fatroy{at}ucdavis.edu.

Abstract

The objective of these studies was to test the hypothesis that proteins which contain potential polyisoprenyl recognition sequences (PIRS) in their transmembrane-spanning domain can bind to the polyisoprenyl (PI) glycosyl carrier lipids, undecaprenylphosphate (C55-P) and dolichylphosphate (C95-P). A number of prokaryotic and eukaryotic glycosyltransferases that utilize PI co-enzymes contain a conserved PIRS postulated to be the active PI binding domain. To study this problem, we first determined the three-dimensional structure of a PIRS peptide, NeuE, by homonuclear 2-D 1H-NMR spectroscopy. Experimentally generated distance constraints derived from NOE and torsion angle constraints derived from coupling constants were used for restrained molecular dynamics and energy minimization calculations. Molecular models of the NeuE peptide were built based on calculations of energy minimization using the DGII program (NMRchitect.). 3-D models of dolichol (C95) and C95-P were built based on our 2-D 1H-NMR NOESY results and refined by energy minimization with respect to all atoms using the AMBER (Assisted Modeling with Energy Refinements) force field. Our energy minimization studies were carried out on a conformational model of dolichol that was originally derived from small-angle X-ray scattering (SAXS) and molecular mechanics methods. These results revealed that the PIs are conformationally nearly identical tripartite molecules, with their three domains arranged in a coiled, helical structure. Analyses of the intermolecular cross peaks in the 2D-NOESY spectra of PIRS peptides in the presence of PIs confirmed a highly specific interaction, and identified key contact amino acids in the NeuE peptide that constituted a "binding motif" for interacting with the PIs. These studies also showed that subtle conformational changes occurred within both the PIs and the NeuE peptide after binding. 3-D structures of the resulting molecular complexes revealed that each PI could bind more than one PIRS peptide. These studies thus represent the first evidence for a direct physical interaction between specific contact amino acids in the PIRS peptides and the PIs, and supports the hypothesis of a bifunctional role for the PIs. The central idea is that these "super-lipids" may serve as a structural scaffold to organize and stabilize in functional domains PIRS-containing proteins within multiglycosyltransferase complexes that participate in biosynthetic and translocation processes.


Key words: glycosyl carrier lipid (dolichol)/conformation-structure of polyisoprenyl recognition sequence peptides/ glycosyl translocation/ NMR
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. Bacteriol.Home page
J. W. Schertzer and E. D. Brown
Use of CDP-Glycerol as an Alternate Acceptor for the Teichoic Acid Polymerase Reveals that Membrane Association Regulates Polymer Length
J. Bacteriol., November 1, 2008; 190(21): 6940 - 6947.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Touze, D. Blanot, and D. Mengin-Lecreulx
Substrate Specificity and Membrane Topology of Escherichia coli PgpB, an Undecaprenyl Pyrophosphate Phosphatase
J. Biol. Chem., June 13, 2008; 283(24): 16573 - 16583.
[Abstract] [Full Text] [PDF]

Home page
 Microbiol. Mol. Biol. Rev.Home page
J. van Heijenoort
Lipid Intermediates in the Biosynthesis of Bacterial Peptidoglycan
Microbiol. Mol. Biol. Rev., December 1, 2007; 71(4): 620 - 635.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
E. Lamani, R. B. Mewbourne, D. S. Fletcher, S. D. Maltsev, L. L. Danilov, V. V. Veselovsky, A. V. Lozanova, N. Ya. Grigorieva, O. A. Pinsker, J. Xing, et al.
Structural studies and mechanism of Saccharomyces cerevisiae dolichyl-phosphate-mannose synthase: insights into the initial step of synthesis of dolichyl-phosphate-linked oligosaccharide chains in membranes of endoplasmic reticulum
Glycobiology, July 1, 2006; 16(7): 666 - 678.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
G.-P. Zhou and F. A. Troy II
NMR study of the preferred membrane orientation of polyisoprenols (dolichol) and the impact of their complex with polyisoprenyl recognition sequence peptides on membrane structure
Glycobiology, April 1, 2005; 15(4): 347 - 359.
[Abstract] [Full Text] [PDF]

Home page
 Microbiol. Mol. Biol. Rev.Home page
E. R. Vimr, K. A. Kalivoda, E. L. Deszo, and S. M. Steenbergen
Diversity of Microbial Sialic Acid Metabolism
Microbiol. Mol. Biol. Rev., March 1, 2004; 68(1): 132 - 153.
[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.