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

Abstract

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.

Authors:
; ; ; ;
Publication Date:
Research Org.:
New York Medical College, Valhalla
OSTI Identifier:
6986156
Report Number(s):
CONF-8606151-
Journal ID: CODEN: FEPRA; TRN: 86-039028
Resource Type:
Conference
Journal Name:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
Additional Journal Information:
Journal Volume: 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
Subject:
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

Citation Formats

Tsukada, H, Carter, S R, Zielenski, J, Slomiany, B L, and Slomiany, A. Studies on the attachment of fatty acid and N-acetylgalactosamine to mucus glycoprotein nascent chains in rat gastric mucosa. United States: N. p., 1986. Web.
Tsukada, H, Carter, S R, Zielenski, J, Slomiany, B L, & Slomiany, A. Studies on the attachment of fatty acid and N-acetylgalactosamine to mucus glycoprotein nascent chains in rat gastric mucosa. United States.
Tsukada, H, Carter, S R, Zielenski, J, Slomiany, B L, and Slomiany, A. 1986. "Studies on the attachment of fatty acid and N-acetylgalactosamine to mucus glycoprotein nascent chains in rat gastric mucosa". United States.
@article{osti_6986156,
title = {Studies on the attachment of fatty acid and N-acetylgalactosamine to mucus glycoprotein nascent chains in rat gastric mucosa},
author = {Tsukada, H and Carter, S R and Zielenski, J and Slomiany, B L and Slomiany, A},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/6986156}, journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 45:6,
place = {United States},
year = {Thu May 01 00:00:00 EDT 1986},
month = {Thu May 01 00:00:00 EDT 1986}
}

Conference:
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