Perturbation of N-linked oligosaccharide structure results in an altered incorporation of (/sup 3/H)palmitate into specific proteins in Chinese hamster ovary cells

Journal Article · Fri Sep 25 00:00:00 EDT 1987 · J. Biol. Chem.; (United States)

OSTI ID:5842192

Wellner, R B; Ghosh, P C; Roecklein, B; Wu, H C

Increased (/sup 3/H)palmitate incorporation into specific cellular proteins has been reported to occur in Chinese hamster ovary and yeast mutant cells. In this paper we report studies concerning the relationship between N-linked oligosaccharide structure and (/sup 3/H)palmitate incorporation into proteins of Chinese hamster ovary (CHO) cells. We have compared the incorporation of (/sup 3/H)palmitate into proteins of wild-type and four different mutant CHO cell lines defective in various steps of N-linked protein glycosylation. Sodium dodecyl sulfate-gel electrophoretic analysis showed that three of the mutants exhibited increased (/sup 3/H)palmitate incorporation into several CHO cellular proteins (approximately 30,000-38,000 molecular weight) as compared to the wild-type cells. One of the affected mutants which accumulates the Man5Gn2Asn intermediate structure was examined in detail. In agreement with earlier reports, virtually all of the (/sup 3/H) palmitate-labeled proteins of both wild-type and mutant cell lines are membrane-bound. Pretreatment of the mutant cell line with tunicamycin blocked the increased (/sup 3/H)palmitate incorporation into the two specific proteins (both of approximately 30,000 molecular weight) observed in untreated cells; the decreased incorporation of (/sup 3/H)palmitate into the 30,000 molecular weight species was accompanied by a concomitant increase in the incorporation of (/sup 3/H)palmitate into two proteins of approximately 20,000 molecular weight. Pretreatment of wild-type cells with tunicamycin also caused increased (/sup 3/H)palmitate incorporation into the 20,000 molecular weight species.

• Research Organization: Uniformed Services University of the Health Sciences, Bethesda, MD
• OSTI ID: 5842192
• Journal Information: J. Biol. Chem.; (United States), Vol. 262:27
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
HEXADECANOIC ACID
METABOLISM
MEMBRANE PROTEINS
BIOSYNTHESIS
CELL MEMBRANES
CHO CELLS
ELECTROPHORESIS
LECTINS
MOLECULAR WEIGHT
MUTANTS
OLIGOSACCHARIDES
PHENOTYPE
STRUCTURE-ACTIVITY RELATIONSHIPS
TRITIUM COMPOUNDS
ANIMAL CELLS
CARBOHYDRATES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
LABELLED COMPOUNDS
MEMBRANES
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
SACCHARIDES
SYNTHESIS
550201* - Biochemistry- Tracer Techniques