Two-dimensional analysis of proteins secreted by Saccharomyces cerevisiae regenerating protoplasts: a novel approach to study the cell wall.
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 beta-1,3- and beta-1, 6-glucosylation was detected in the proteins secreted by regenerating protoplasts.
Pardo
M
M
Departamento de Microbiología II, Facultad de Farmacia Universidad Complutense, Madrid, Spain.
Monteoliva
L
L
Pla
J
J
Sánchez
M
M
Gil
C
C
Nombela
C
C
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002473:Cell Wall / Q000737:chemistry* / Q000201:enzymology / Q000378:metabolism / Q000502:physiology; D017077:Culture Media, Conditioned; D015180:Electrophoresis, Gel, Two-Dimensional; D005656:Fungal Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005966:Glucuronidase / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000235:genetics; D006031:Glycosylation; D006360:Heat-Shock Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000378:metabolism; D009154:Mutation; D011523:Protoplasts / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D017421:Sequence Analysis; D013395:Sucrose / Q000378:metabolism; D013429:Sulfatases / Q000378:metabolism; D043324:beta-Fructofuranosidase
D017077:Culture Media, Conditioned; D005656:Fungal Proteins; D006360:Heat-Shock Proteins; D009097:Multienzyme Complexes; C023059:glusulase; D013395:Sucrose; D013429:Sulfatases; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D005966:Glucuronidase