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Microbiology 149 (2003), 2769-2778; DOI  10.1099/mic.0.26527-0
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Microbiology 149 (2003), 2769-2778; DOI  10.1099/mic.0.26527-0
© 2003 Society for General Microbiology

Identification and characterization of a novel autolysin (Aae) with adhesive properties from Staphylococcus epidermidis

Christine Heilmann1, Günther Thumm2, Gursharan S. Chhatwal3, Jörg Hartleib1, Andreas Uekötter1 and Georg Peters1

1 Institute of Medical Microbiology, University of Münster, Domagkstr. 10, D-48149 Münster, Germany
2 Mikrobielle Genetik, University of Tübingen, Auf der Morgenstelle 28, D-72076 Tübingen, Germany
3 Division of Microbiology, GBF-National Research Center for Biotechnology, Spielmannstr. 7, D-38106 Braunschweig, Germany

Correspondence
Christine Heilmann
heilmac{at}uni-muenster.de

Staphylococcus epidermidis biofilm formation on polymer surfaces is considered a major pathogenicity factor in foreign-body-associated infections. Previously, the 148 kDa autolysin AtlE from S. epidermidis, which is involved in the initial attachment of the cells to polymer surfaces and also binds to the extracellular matrix protein vitronectin, was characterized. Here, the characterization of a novel autolysin/adhesin (Aae) in S. epidermidis is described. Aae was identified as a 35 kDa surface-associated protein that has bacteriolytic activity and binds vitronectin. Its N-terminal amino acid sequence was determined and the respective gene, aae, was cloned. DNA-sequence analysis revealed that aae encodes a deduced protein of 324 amino acids with a predicted molecular mass of 35 kDa. Aae contains three repetitive sequences in its N-terminal portion. These repeats comprise features of a putative peptidoglycan binding domain (LysM domain) found in a number of enzymes involved in cell-wall metabolism and also in some adhesins. Expression of aae by Escherichia coli and subsequent analysis revealed that Aae possesses bacteriolytic activity and adhesive properties. The interaction of Aae with fibrinogen, fibronectin and vitronectin was found to be dose-dependent and saturable and to occur with high affinity, by using the real-time Biomolecular Interaction Analysis (BIA). Aae binds to the A{alpha}- and B{beta}-chains of fibrinogen and to the 29 kDa N-terminal fragment of fibronectin. In conclusion, Aae is a surface-associated protein with bacteriolytic and adhesive properties representing a new member of the staphylococcal autolysin/adhesins potentially involved in colonization.


Abbreviations: Aae, autolysin/adhesin from S. epidermidis; BIA, Biomolecular Interaction Analysis; Fg, fibrinogen; Fn, fibronectin; His6, hexahistidine; Vn, vitronectin

The EMBL/GenBank/DDBJ accession number for the aae DNA sequence reported in this article is AJ250905.




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