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A proteomic approach for the study of Saccharomyces cerevisiae cell wall biogenesis
Author(s) -
Pardo Mercedes,
Ward Malcolm,
Bains Satty,
Molina María,
Blackstock Walter,
Gil Concha,
Nombela César
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20001001)21:16<3396::aid-elps3396>3.0.co;2-j
Subject(s) - cell wall , biology , biogenesis , extracellular , cell , proteomics , microbiology and biotechnology , biochemistry , protoplast , secretion , gel electrophoresis , proteome , secretory protein , gene
In fungi, cell shape is determined by the presence of a rigid cell wall which separates the cell from the extracellular medium. This highly dynamic structure is essential for the maintenance of cell integrity and is involved in several phenomena such as flocculation, adherence and pathogenicity. The composition of the fungal cell wall is well known, but issues such as the assembly and remodeling of its components remain poorly understood. In an attempt to study the de novo construction of the yeast cell wall, we have undertaken a large‐scale proteomic approach to analyze the proteins secreted by regenerating protoplasts. Upon incubation of protoplasts in regenerating conditions, numerous proteins are secreted into the culture medium. These presumably include proteins destined for the cell wall, comprising both structural proteins as well as enzymes involved in cell wall biogenesis. This work reports the establishment of a reference map of proteins secreted by regenerating protoplasts by means of two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) and their identification by mass spectrometry. Thirty‐two different proteins have been identified, including known cell wall proteins, glycolytic enzymes, heat shock proteins, and proteins involved in several other processes. Using this approach, novel proteins possibly involved in cell wall construction have also been identified. This reference map will allow comparative analyses to be carried out on a selected collection of mutants affected in the cell wall.