Abstract
Cells from the “slime” variant of Neurospora crassa were broken in isotonic conditions by use of triethanolamine buffer plus EDTA. After removal of large membranous structures by low-speed centrifugation, chitosomes and secretory vesicles were separated by means of gel filtration, precipitation of membranous contaminants with Concanavalin A, and centrifugation in sucrose or glycerol gradients. Polypeptidic composition of fractions enriched in secretory vesicles or chitosomes was found to be distinct. By these criteria we concluded that chitosomes and secretory vesicles represent different populations of microvesicles. Both microvesicular populations appeared free of endoplasmic reticulum and vacuolar contaminants as demonstrated by determination of appropriate enzymatic markers.
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Abbreviations
- ER:
-
Endoplasmic reticulum
- UDP-GlcNAc:
-
uridine-5′-diphosphate N-acetyl glucosamine
- GlcNAc:
-
N-acetyl glucosamine
- SDS:
-
sodium dodecyl sulfate
- PMSF:
-
phenyl methyl sulfonyl fluoride
- EDTA:
-
ethylene diamino tetraacetic acid
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Investigador Nacional de Mexico. On leave from the Centro de Investigacion y Estudios Avanzados (IPN), and the Universidad de Guanajuato, Gto., Mexico
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Ruiz-Herrera, J., Martinez, J.P., Casanova, M. et al. Separation of chitosomes and secretory vesicles from the “slime” variant of Neurospora crassa . Arch. Microbiol. 149, 156–162 (1987). https://doi.org/10.1007/BF00425082
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DOI: https://doi.org/10.1007/BF00425082