Premium
Two‐Dimensional analysis of proteins secreted by Saccharomyces cerevisiae regenerating protoplasts: a novel approach to study the cell wall
Author(s) -
Pardo Mercedes,
Monteoliva Lucía,
Pla Jesús,
Sánchez Miguel,
Gil Concha,
Nombela César
Publication year - 1999
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(199904)15:6<459::aid-yea387>3.0.co;2-l
Subject(s) - biology , protoplast , cell wall , saccharomyces cerevisiae , biochemistry , secretory protein , secretion , enzyme , heat shock protein , cell , two dimensional gel electrophoresis , yeast , microbiology and biotechnology , gene , proteomics
Protoplasts of Saccharomyces cerevisiae incubated in regenerating conditions secrete cell wall components in order to allow the biosynthesis of this structure. During the first hours of incubation, many of these are not retained in the forming cell wall but remain in the medium. We have developed a method for collecting the secreted proteins and have analysed these by two‐dimensional electrophoresis to obtain a reference map of putative cell wall proteins. Several proteins were identified by microsequencing or immunoblotting; namely, cell wall hydrolytic enzymes, heat shock proteins, glycolytic enzymes and others. Some β‐1,3‐ and β‐1,6‐glucosylation was detected in the proteins secreted by regenerating protoplasts. Copyright © 1999 John Wiley & Sons, Ltd.