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Phosphatidylinositol-anchored glycoproteins of PC12 pheochromocytoma cells and brain

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00409a049· OSTI ID:7000175
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PC12 pheochromocytoma cells and cultures of early postnatal rat cerebellium were labeled with (/sup 3/H)glucosamine, (/sup 3/H)fucose, (/sup 3/H)leucine, (/sup 3/H)ethanolamine, or sodium (/sup 35/S)sulfate and treated with a phosphatidylinositol-specific phospholipase C. Enzyme treatment of (/sup 3/H) glucosamine- or (/sup 3/H)fucose-labeled PC12 cells led to a 15-fold increase in released glycoproteins. On sodium dodecyl sulfate-polyacrylamide gel ectrophoresis, most of the released material migrated as a broad band with an apparent molecular size of 32,000 daltons (Da), which was specifically immunoprecipitated by a monoclonal antibody to the Thy-l glycoprotein. A second glycoprotein, with an apparent molecular size of 158,000 Da, was also released. After treatment with endo-..beta..-galactosidase, 40-45% of the (/sup 3/H)glucosamine of (/sup 3/H)fucose radioactivity in the phospholipase-released glycoproteins was converted to products of disaccharide size, and the molecular size of the 158-kDa glycoprotein decreased to 145 kDa, demonstrating that it contains fucosylated poly-(N-acetyllactosaminyl) oligosaccharides. The phospholipase also released labeled Thy-1 and the 158-kDa glycoprotein from PC12 cells cultured in the presence of (/sup 3/H)ethanolamine, which specifically labels this component of the phosphatidylinositol membrane-anchoring sequence,while in the lipid-free protein residue of cells not treated with phospholipase, Thy-1 and a doublet at 46/48 kDa were the only labeled proteins. Sulfated glycoproteins of 155, 132/134, 61, and 21 kDa are the predominant species released by phospholipase, which does not affect a major 44-kDa protein seen in (/sup 3/H)ethanolamine-labeled brain cultures. The 44-48- and 155/158-kDa proteins may be common to both PC12 cells and brain.

Research Organization:
State Univ. of New York Health Science Center, Brooklyn (USA)
OSTI ID:
7000175
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 27:9; ISSN BICHA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALI METAL COMPOUNDS
AMINES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BODY
BRAIN
CARBOHYDRATES
CARBOXYLIC ACIDS
CELL CULTURES
CENTRAL NERVOUS SYSTEM
CHEMISTRY
DAYS LIVING RADIOISOTOPES
ELECTROPHORESIS
EVEN-ODD NUCLEI
GLUCOPROTEINS
GLUCOSAMINE
HEXOSAMINES
HEXOSES
ISOTOPES
LABELLED COMPOUNDS
LEUCINE
LIGHT NUCLEI
MAMMALS
MOLECULAR STRUCTURE
MONOSACCHARIDES
NERVOUS SYSTEM
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SACCHARIDES
SODIUM COMPOUNDS
SODIUM SULFATES
SULFATES
SULFUR 35
SULFUR COMPOUNDS
SULFUR ISOTOPES
TRITIUM COMPOUNDS
TUMOR CELLS
VERTEBRATES