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Infect. Immun. doi:10.1128/IAI.01049-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

-Enolase resides on the cell surface of Mycoplasma fermentans and binds plasminogen

Department of Membrane and Ultrastructure Research, The Hebrew University-Hadassah Medical School, Jerusalem, Israel

^* To whom correspondence should be addressed. Email: Rottem{at}cc.huji.ac.il.

	Abstract

Plasminogen (Plg) binding to the cell surface of Mycoplasma fermentans resulted in a marked increase in the maximal adherence of the organism to HeLa cells, an enhanced Plg activation by the urokinase-type Plg activator and in the induction of the internalization of M. fermentans by host eukaryotic cells (Yavlovich, A., A. Katzenell, M. Tarshis, A. A. Higazi and S. Rottem. Mycoplasma fermentans binds to and invades HeLa cells: involvement of plasminogen and urokinase. Infect. Immun. 72:5004-5011, 2004). In this study the M. fermentans Plg-binding protein was isolated by affinity chromatography of Triton X-100 solubilized M. fermentans membranes utilizing a column of Plg-biotin complex attached to avidin that was eluted with -amino caproic acid. The eluted 50 kDa protein was identified by mass spectrometric techniques as -enolase. The possibility that -enolase, a key cytoplasmatic glycolytic enzyme, resides also on the cell surface of M. fermentans was supported by immunoblot analysis using polyclonal anti -enolase antiserum showing that -enolase is present in a purified M. fermentans membrane preparation, as well as by immunochemical criteria and by immuno-electron microscopy analysis. Our observation that Plg blocked binding of anti -enolase antibodies to a 50 kDa polypeptide band resolved by SDS-PAGE of M. fermentans membrane or soluble preparations further supports our notion that mycoplasmal surface -enolase is a major Plg binding protein of M. fermentans.