Characteristics of oligosaccharides from rat parotid (RP) N-linked glycoproteins (GP) after. beta. -adrenoreceptor (. beta. -AR) stimulation

Conference · Fri May 01 04:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6024280

Baum, B J; Yeh, C K; Kousvelari, E E

The authors have shown that ..beta..-AR stimulation of RP cells leads to marked enhancement of N-linked glycosylation in 4 secretory GP (Mrapprox.220Kd, HMW;approx.32-38Kd, MMW;approx.17Kd, LMW). To characterize oligosaccharides in GP, cells were incubated 60 min +/- isoproterenol (ISO) and analyzed 2 ways. First, cell extracts were subjected to SDS-PAGE and Western blotting with peroxidase-conjugated Con A or wheat germ agglutinin (WGA). Second, double-labeled (/sup 3/H) man//sup 14/C leu) extracts were chromatographed on G200 followed by analysis of GP on Con A-Sepharose. HMW from control (CON) and ISO cells were Endo H insensitive, Endo F sensitive, altered by incubation with deoxynojirimycin (dNM), and bound both Con A and WGA conjugates. Similar findings were observed with LMW while MMW were sensitive to Endo H and Endo F, unaffected by dNM, bound Con A (strongly) and WGA (weakly). MMW and LMW were primarily eluted from Con A-Sepharose with 0.5M ..cap alpha..-methyl mannoside (..cap alpha..-MM) while HMW were eluted sequentially with 10 mM ..cap alpha..-methyl glucoside and ..cap alpha..-MM. HMW, MMW, and LMW had approx.4 fold higher /sup 3/H//sup 14/C ratios after ISO. These results suggest HMW and LMW likely contain biantennary complex and hybrid oligosaccharides while MMW contain only high mannose oligosaccharide types.

• Research Organization: National Institutes of Health, Bethesda, MD
• OSTI ID: 6024280
• Report Number(s): CONF-870644-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
BIOCHEMICAL REACTION KINETICS
BODY
CARBOHYDRATES
CARBON 14 COMPOUNDS
CHEMICAL PROPERTIES
CHROMATOGRAPHY
DOUBLE LABELLING
DRUGS
ELECTROPHORESIS
GLANDS
GLUCOPROTEINS
ISOTOPE APPLICATIONS
ISOTOPE RATIO
KINETICS
LABELLED COMPOUNDS
LABELLING
MAMMALS
MEMBRANE PROTEINS
METABOLISM
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RATS
REACTION KINETICS
RECEPTORS
RODENTS
SACCHARIDES
SALIVARY GLANDS
SEPARATION PROCESSES
SYMPATHOMIMETICS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES