Alpha-enolase resides on the cell surface of Mycoplasma fermentans and binds plasminogen.
Plasminogen (Plg) binding to the cell surface of Mycoplasma fermentans results in a marked increase in the maximal adherence of the organism to HeLa cells, enhanced Plg activation by the urokinase-type Plg activator, and the induction of the internalization of M. fermentans by eukaryotic host cells (A. Yavlovich, A. Katzenell, M. Tarshis, A. A. Higazi, and S. Rottem, 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 by utilizing a column of a Plg-biotin complex attached to avidin that was eluted with epsilon-aminocaproic acid. The eluted approximately 50-kDa protein was identified by mass spectrometric techniques as alpha-enolase. The possibility that alpha-enolase, a key cytoplasmatic glycolytic enzyme, resides also on the cell surface of M. fermentans was supported by an immunoblot analysis using polyclonal anti-alpha-enolase antiserum, which showed that alpha-enolase was present in a purified M. fermentans membrane preparation, as well as by immunochemical criteria and by immunoelectron microscopy analysis. Our observation that Plg blocked the binding of anti-alpha-enolase antibodies to a 50-kDa polypeptide band resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of M. fermentans membrane or soluble preparations further supports our notion that mycoplasmal surface alpha-enolase is a major Plg binding protein of M. fermentans.
Yavlovich
Amichai
A
Department of Membrane and Ultrastructure Research, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
Rechnitzer
Hagai
H
Rottem
Shlomo
S
D002846:Chromatography, Affinity / Q000379:methods; D015151:Immunoblotting / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins / Q000378:metabolism; D016253:Microscopy, Immunoelectron / Q000379:methods; D009175:Mycoplasma Infections / Q000097:blood / Q000378:metabolism* / Q000382:microbiology; D016858:Mycoplasma fermentans / Q000201:enzymology* / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism*; D010958:Plasminogen / Q000276:immunology / Q000378:metabolism*
D008565:Membrane Proteins; D010958:Plasminogen; D010751:Phosphopyruvate Hydratase