Isolation of secretory vesicles from Saccharomyces cerevisiae.
Purification of secretory vesicles from Saccharomyces cerevisiae has been hindered because these organelles normally represent a small proportion of cellular membranes. In the yeast secretory mutant sec1, secretory vesicles accumulate intracellularly in large quantities. Using a sec1 strain we have devised a procedure for the partial purification of these vesicles. The purification employs differential and density gradient centrifugations and an electrophoretic separation of membranes. The fractions obtained from this procedure are enriched for secretory vesicles at least fivefold over other cellular membranes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of solubilized membrane fractions reveals a distinct set of polypeptides associated with secretory vesicles.
Holcomb
C L
CL
Department of Biochemistry, University of California, Berkeley 94720.
Etcheverry
T
T
Schekman
R
R
D000135:Acid Phosphatase / Q000302:isolation & purification; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000648:ultrastructure*; D012967:Sodium Dodecyl Sulfate; D013056:Spectrophotometry, Ultraviolet; D013104:Spheroplasts / Q000648:ultrastructure; D013462:Sulfur Radioisotopes
D005656:Fungal Proteins; D013462:Sulfur Radioisotopes; D012967:Sodium Dodecyl Sulfate; D000135:Acid Phosphatase