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Journal of Bacteriology, August 2004, p. 4931-4939, Vol. 186, No. 15
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.15.4931-4939.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Fibronectin Binding to the Salmonella enterica Serotype Typhimurium ShdA Autotransporter Protein Is Inhibited by a Monoclonal Antibody Recognizing the A3 Repeat

Robert A. Kingsley,1* Daad Abi Ghanem,2 Nahum Puebla-Osorio,2 A. Marijke Keestra,1 Luc Berghman,2 and Andreas J. Bäumler1

Department of Medical Microbiology and Immunology, College of Medicine, College Station, Texas 77843-1114,1 Departments of Poultry Science and Veterinary Pathobiology, Texas A&M University, College Station, Texas 77843-24722

Received 3 February 2004/ Accepted 26 April 2004

ShdA is a large outer membrane protein of the autotransporter family whose passenger domain binds the extracellular matrix proteins fibronectin and collagen I, possibly by mimicking the host ligand heparin. The ShdA passenger domain consists of ~1,500 amino acid residues that can be divided into two regions based on features of the primary amino acid sequence: an N-terminal nonrepeat region followed by a repeat region composed of two types of imperfect direct amino acid repeats, called type A and type B. The repeat region bound bovine fibronectin with an affinity similar to that for the complete ShdA passenger domain, while the nonrepeat region exhibited comparatively low fibronectin-binding activity. A number of fusion proteins containing truncated fragments of the repeat region did not bind bovine fibronectin. However, binding of the passenger domain to fibronectin was inhibited in the presence of immune serum raised to one truncated fragment of the repeat region that contained repeats A2, B8, A3, and B9. Furthermore, a monoclonal antibody that specifically recognized an epitope in a recombinant protein containing the A3 repeat inhibited binding of ShdA to fibronectin.

* Corresponding author. Present address: The Wellcome Trust Sanger Institute, The Wellcome Trust Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, United Kingdom. Phone: 44 (0)1223 495381. Fax: 44 (0)1223 494919. E-mail: rak{at}sanger.ac.uk.

Journal of Bacteriology, August 2004, p. 4931-4939, Vol. 186, No. 15
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.15.4931-4939.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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