HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 30, 30945-30953, July 23, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/30/30945    most recent M401613200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Daubenspeck, J. M. Articles by Turnbough, C. L. Search for Related Content PubMed PubMed Citation Articles by Daubenspeck, J. M. Articles by Turnbough, C. L., Jr. Social Bookmarking                      What's this?

Novel Oligosaccharide Side Chains of the Collagen-like Region of BclA, the Major Glycoprotein of the Bacillus anthracis Exosporium^*

James M. Daubenspeck, Huadong Zeng, Ping Chen, Shengli Dong¶, Christopher T. Steichen, N. Rama Krishna¶, David G. Pritchard¶, and Charles L. Turnbough, Jr.||

From the Department of Microbiology, Comprehensive Cancer Center, and ^¶Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294

Spores of Bacillus anthracis, the causative agent of anthrax, are enclosed by a prominent loose fitting layer called the exosporium. The exosporium consists of a basal layer and an external hairlike nap. The filaments of the nap are composed of a highly immunogenic glycoprotein called BclA, which has a long, central collagen-like region with multiple XXG repeats. Most of the triplet repeats are PTG, and nearly all of the triplet repeats contain a threonine residue, providing multiple potential sites for O-glycosylation. In this study, we demonstrated that two O-linked oligosaccharides, a 715-Da tetrasaccharide and a 324-Da disaccharide, are released from spore- and exosporium-associated BclA by hydrazinolysis. Each oligosaccharide is probably attached to BclA through a GalNAc linker, which was lost during oligosaccharide release. We found that multiple copies of the tetrasaccharide are linked to the collagen-like region of BclA, whereas the disaccharide may be attached outside of this region. Using NMR, mass spectrometry, and other analytical techniques, we determined that the structure of the tetrasaccharide is 2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy--D-glucopyranosyl-(13)--L-rhamnopyranosyl-(13)--L-rhamnopyranosyl-(12)-L-rhamnopyranose. The previously undescribed nonreducing terminal sugar (i.e. 2-O-methyl-4-(3-hydroxy-3-methylbutamido)-4,6-dideoxy-D-glucose) was given the trivial name anthrose. Anthrose was not found in spores of either Bacillus cereus or Bacillus thuringiensis, two species that are the most phylogenetically similar to B. anthracis. Thus, anthrose may be useful for species-specific detection of B. anthracis spores or as a new target for therapeutic intervention.

Received for publication, February 13, 2004 , and in revised form, May 17, 2004.

^* This work was supported by National Institutes of Health (NIH) Grant AI50566. The University of Alabama at Birmingham NMR Core Facility was supported by NCI, NIH, Grant CA13148 and National Science Foundation Instrumentation Grant DBI-9729461. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Dept. of Microbiology, University of Alabama at Birmingham, BBRB 409, 1530 3rd Ave. S., Birmingham, AL 35294. Tel.: 205-934-6289; Fax: 205-975-5479; E-mail: ChuckT{at}uab.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. Leoff, E. Saile, J. Rauvolfova, C. P Quinn, A. R Hoffmaster, W. Zhong, A. S Mehta, G.-J. Boons, R. W Carlson, and E. L Kannenberg Secondary cell wall polysaccharides of Bacillus anthracis are antigens that contain specific epitopes which cross-react with three pathogenic Bacillus cereus strains that caused severe disease, and other epitopes common to all the Bacillus cereus strains tested Glycobiology, June 1, 2009; 19(6): 665 - 673. [Abstract] [Full Text] [PDF]

				J. Kubler-Kielb, E. Vinogradov, H. Hu, S. H. Leppla, J. B. Robbins, and R. Schneerson Saccharides cross-reactive with Bacillus anthracis spore glycoprotein as an anthrax vaccine component PNAS, June 24, 2008; 105(25): 8709 - 8712. [Abstract] [Full Text] [PDF]

				S. Dong, S. A. McPherson, L. Tan, O. N. Chesnokova, C. L. Turnbough Jr., and D. G. Pritchard Anthrose Biosynthetic Operon of Bacillus anthracis J. Bacteriol., April 1, 2008; 190(7): 2350 - 2359. [Abstract] [Full Text] [PDF]

				C. Leoff, E. Saile, D. Sue, P. Wilkins, C. P. Quinn, R. W. Carlson, and E. L. Kannenberg Cell Wall Carbohydrate Compositions of Strains from the Bacillus cereus Group of Species Correlate with Phylogenetic Relatedness J. Bacteriol., January 1, 2008; 190(1): 112 - 121. [Abstract] [Full Text] [PDF]

				A. Mohs, T. Silva, T. Yoshida, R. Amin, S. Lukomski, M. Inouye, and B. Brodsky Mechanism of Stabilization of a Bacterial Collagen Triple Helix in the Absence of Hydroxyproline J. Biol. Chem., October 12, 2007; 282(41): 29757 - 29765. [Abstract] [Full Text] [PDF]

				K. Takeuchi, H. Ono, M. Yoshida, T. Ishii, E. Katoh, F. Taguchi, R. Miki, K. Murata, H. Kaku, and Y. Ichinose Flagellin Glycans from Two Pathovars of Pseudomonas syringae Contain Rhamnose in D and L Configurations in Different Ratios and Modified 4-Amino-4,6-Dideoxyglucose J. Bacteriol., October 1, 2007; 189(19): 6945 - 6956. [Abstract] [Full Text] [PDF]

				B. M. Thompson, L. N. Waller, K. F. Fox, A. Fox, and G. C. Stewart The BclB Glycoprotein of Bacillus anthracis Is Involved in Exosporium Integrity J. Bacteriol., September 15, 2007; 189(18): 6704 - 6713. [Abstract] [Full Text] [PDF]

				J. A. Boydston, L. Yue, J. F. Kearney, and C. L. Turnbough Jr. The ExsY Protein Is Required for Complete Formation of the Exosporium of Bacillus anthracis J. Bacteriol., November 1, 2006; 188(21): 7440 - 7448. [Abstract] [Full Text] [PDF]

				M. K. Swiecki, M. W. Lisanby, F. Shu, C. L. Turnbough Jr, and J. F. Kearney Monoclonal Antibodies for Bacillus anthracis Spore Detection and Functional Analyses of Spore Germination and Outgrowth J. Immunol., May 15, 2006; 176(10): 6076 - 6084. [Abstract] [Full Text] [PDF]

				S. Rety, S. Salamitou, I. Garcia-Verdugo, D. J. S. Hulmes, F. Le Hegarat, R. Chaby, and A. Lewit-Bentley The Crystal Structure of the Bacillus anthracis Spore Surface Protein BclA Shows Remarkable Similarity to Mammalian Proteins J. Biol. Chem., December 30, 2005; 280(52): 43073 - 43078. [Abstract] [Full Text] [PDF]

				C. T. Steichen, J. F. Kearney, and C. L. Turnbough Jr Characterization of the Exosporium Basal Layer Protein BxpB of Bacillus anthracis J. Bacteriol., September 1, 2005; 187(17): 5868 - 5876. [Abstract] [Full Text] [PDF]

				P. Sylvestre, E. Couture-Tosi, and M. Mock Contribution of ExsFA and ExsFB Proteins to the Localization of BclA on the Spore Surface and to the Stability of the Bacillus anthracis Exosporium J. Bacteriol., August 1, 2005; 187(15): 5122 - 5128. [Abstract] [Full Text] [PDF]

				J. A. Boydston, P. Chen, C. T. Steichen, and C. L. Turnbough Jr. Orientation within the Exosporium and Structural Stability of the Collagen-Like Glycoprotein BclA of Bacillus anthracis J. Bacteriol., August 1, 2005; 187(15): 5310 - 5317. [Abstract] [Full Text] [PDF]

				L. N. Waller, M. J. Stump, K. F. Fox, W. M. Harley, A. Fox, G. C. Stewart, and M. Shahgholi Identification of a Second Collagen-Like Glycoprotein Produced by Bacillus anthracis and Demonstration of Associated Spore-Specific Sugars J. Bacteriol., July 1, 2005; 187(13): 4592 - 4597. [Abstract] [Full Text] [PDF]