Enolase is present in the cell wall of Saccharomyces cerevisiae

Non-covalently attached or soluble cell wall proteins of Saccharomyces cere visiae were extracted using a high pH/2-mercaptoethanol procedure and were separated for peptide sequencing using 2D-PAGE. A partial N- terminal seque nce of a major protein spot was obtained and showed high identity with enol ase gene products. Western blotting with an anti-enolase antibody confirmed that enolase was present in the cell wall extract. Biotinylation of cells prior to protein extraction with a membrane impermeable biotinylating agent confirmed that the detection was not owing to cell lysis during extraction . Transmission immunoelectron microscopy showed enolase to be present in th e cell wall. Enolase contains no known secretion signal that would localize it to the cell wall. Thus S. cevevisiae must have further mechanisms for t argeting proteins to the cell wall. (C) 1999 Federation of European Microbi ological Societies. Published by Elsevier Science B.V. All rights reserved.