Studies on the attachment of fatty acid and N-acetylgalactosamine to mucus glycoprotein nascent chains in rat gastric mucosa

Conference · Thu May 01 00:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6986156

Tsukada, H; Carter, S R; Zielenski, J; Slomiany, B L; Slomiany, A

The authors have investigated the process of mucin protein acylation and glycosylation to ascertain whether fatty acids and galactosamine are added to nascent chains prior to the completion or after release from the ribosome. For fatty acid and carbohydrate labeling, the rat gastric mucosal cell suspension was incubated with (/sup 3/H) palmitic acid, and (/sup 14/C)-N-acetylgalactosamine for 3h at 37/sup 0/C. After incubation, the cells were washed with incubation medium and subjected to subcellular fractionation. The peptidyl-tRNA was released from polysomes by sedimentation in sucrose gradient containing EDTA (20-30% sucrose gradient). The fraction containing peptidyl-tRNA was dialyzed and nascent chains were separated by ion-exchange chromatography on DEAE-Sephadex. The peptidyl-tRNA, eluted with a linear NaCl gradient was hydrolyzed and separated by SDS-gel electrophoresis and thin-layer chromatography. The individual bands isolated from thin layer plates were analyzed for the content of (/sup 3/H) palmitic acid and (/sup 14/C) N-acetylgalactosamine. The results revealed that fatty acids are incorporated into nascent chains prior to chain completion. Gel electrophoresis and thin layer chromatography showed that the fatty acid moiety is added soon after the acylation site is synthesized on the ribosome, while the GalNAc becomes incorporated only to the acylated peptide.

• Research Organization: New York Medical College, Valhalla
• OSTI ID: 6986156
• Report Number(s): CONF-8606151-; TRN: 86-039028
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CARBOXYLIC ACIDS
BIOCHEMICAL REACTION KINETICS
GLUCOPROTEINS
ACYLATION
OLIGOSACCHARIDES
CARBON 14 COMPOUNDS
ELECTROPHORESIS
HEXADECANOIC ACID
ION EXCHANGE CHROMATOGRAPHY
MUCOUS MEMBRANES
RATS
STOMACH
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TRANSFER RNA
TRITIUM COMPOUNDS
ANIMALS
BODY
CARBOHYDRATES
CHEMICAL REACTIONS
CHROMATOGRAPHY
DIGESTIVE SYSTEM
GASTROINTESTINAL TRACT
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANES
MONOCARBOXYLIC ACIDS
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PROTEINS
REACTION KINETICS
RNA
RODENTS
SACCHARIDES
SEPARATION PROCESSES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques