A proteomic approach for the study of Saccharomyces cerevisiae cell wall biogenesis

Citation
M. Pardo et al., A proteomic approach for the study of Saccharomyces cerevisiae cell wall biogenesis, ELECTROPHOR, 21(16), 2000, pp. 3396-3410
Citations number
85
Categorie Soggetti
Chemistry & Analysis
Journal title
ELECTROPHORESIS
ISSN journal
01730835 → ACNP
Volume
21
Issue
16
Year of publication
2000
Pages
3396 - 3410
Database
ISI
SICI code
0173-0835(200010)21:16<3396:APAFTS>2.0.ZU;2-S
Abstract
In fungi, cell shape is determined by the presence of a rigid cell wall whi ch separates the cell from the extracellular medium. This highly dynamic st ructure 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 a ttempt to study the de novo construction of the yeast cell wall, we have un dertaken 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 str uctural proteins as well as enzymes involved in cell wall biogenesis. This work reports the establishment of a reference map of proteins secreted by r egenerating protoplasts by means of two-dimensional polyacrylamide gel elec trophoresis (2-D PAGE) and their identification by mass spectrometry. Thirt y-two different proteins have been identified, including known cell wall pr oteins, glycolytic enzymes, heat shock proteins, and proteins involved in s everal other processes. Using this approach, novel proteins possibly involv ed 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.