This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
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 arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ryu, H.
Right arrow Articles by Cassels, F. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ryu, H.
Right arrow Articles by Cassels, F. J.

 Previous Article  |  Next Article 

Infection and Immunity, February 2001, p. 640-649, Vol. 69, No. 2
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.2.640-649.2001

Escherichia coli Strain RDEC-1 AF/R1 Endogenous Fimbrial Glycoconjugate Receptor Molecules in Rabbit Small Intestine

Hyoik Ryu,1,dagger Young S. Kim,2 Philippe A. Grange,1 and Frederick J. Cassels1,*

Department of Enteric Infections, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910-7500,1 and Gastrointestinal Research Laboratory, Veterans Administration Medical Center, San Francisco, California 941212

Received 16 March 2000/Returned for modification 8 May 2000/Accepted 26 October 2000

Escherichia coli strain RDEC-1 causes a diarrheagenic infection in rabbits with AF/R1 fimbriae, which have been identified as an important colonization factor in RDEC-1 adherence leading to disease. The AF/R1-mediated RDEC-1 adherence model has been used as a model systems for E. coli diarrheal diseases. In this study, RDEC-1 adhered specifically to small intestinal brush borders, with both sialic acid and beta -galactosyl residues apparently involved. The AF/R1-mediated adherence activity of [14C]-labeled RDEC-1 was analyzed quantitatively by using 24-well plates coated with purified brush borders and purified microvilli. Two microvillus membrane proteins (130 and 140 kDa) were individually isolated, and chicken antibody raised to each protein inhibited bacterial adherence. These same two proteins, previously shown to be recognized by AF/R1, were individually digested with trypsin, and the amino acid sequences of peptides were determined by reversed-phase capillary liquid chromatography-mass spectrometry tandem mass spectrometry (LC-MS). This LC-MS analysis indicated that these proteins are subunits of the rabbit sucrase-isomaltase protein (SI) complex. Guinea pig serum raised to purified rabbit SI complex inhibited bacterial adherence to microvilli. Additionally, as determined by high-performance thin-layer chromatography and autoradiography, RDEC-1 adhered selectively, via AF/R1 fimbriae, to a glycolipid tentatively identified as galactosylceramide (Galbeta 1-1Cer) in the lipid extract of rabbit small intestinal brush borders. RDEC-1 adherence to Galbeta 1-1Cer was partially inhibited in the presence of galactose. These combined results indicate that the endogenous receptor molecule for AF/R1 fimbriae of RDEC-1 is each individual component of the SI complex, although binding to glycolipid may be responsible for an additional adherence mechanism.

* Corresponding author. Mailing address: Department of Enteric Infections, Walter Reed Army Institute of Research, 503 Robert Grant Ave., Silver Spring, MD 20910-7500. Phone: (301) 319-9798. Fax: (301) 319-9801. E-mail: fred.cassels{at}na.amedd.army.mil.

dagger Present address: Department of Biochemistry, Yeungnam University, Kyungsan, 712-749 Republic of Korea.

Infection and Immunity, February 2001, p. 640-649, Vol. 69, No. 2
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.2.640-649.2001


This article has been cited by other articles:

  • McConnell, R. E., Higginbotham, J. N., Shifrin, D. A. Jr., Tabb, D. L., Coffey, R. J., Tyska, M. J. (2009). The enterocyte microvillus is a vesicle-generating organelle. JCB 185: 1285-1298 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»