The relation between glycosylation and activity of guinea pig lipoprotein lipase.
Previous studies have indicated that the processing of oligosaccharide chains is necessary for lipoprotein lipase to become catalytically active and may be involved in the regulation of lipase release. Guinea pig adipocytes and perfused guinea pig hearts were labeled with [35S]methionine, and lipoprotein lipase was immunoprecipitated. Digestion with endo-beta-N-acetylglucosaminidase H (Endo H) showed that the mature enzyme contains one high mannose and two complex oligosaccharide chains. Limited proteolysis indicated where in the molecule the chains are attached. Pulse-chase experiments showed that some lipase molecules were rapidly processed and appeared in the medium within 40 min. Other lipase molecules remained fully Endo H-sensitive for more than 2 h, and this form of the lipase did not appear in the medium. Both forms co-eluted with the sole lipoprotein lipase activity peak from heparin-Sepharose; this indicates that both were dimeric. Separation of the two forms was achieved by lectin chromatography and demonstrated that both were catalytically active. Cells treated with methyl-deoxynojirimycin or with deoxymannojirimycin produced and released active lipoprotein lipase which was fully Endo H-sensitive. These studies demonstrate that the trimming and processing of the oligosaccharide chains is not necessary for lipoprotein lipase to become catalytically active and be secreted, and they suggest that a comparatively large fraction of the lipase molecules is retained in the endoplasmic reticulum. Whether they ever reach the processing apparatus in the Golgi or are degraded is not clear.
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Department of Physiological Chemistry, University of Umeå, Sweden.
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D017485:1-Deoxynojirimycin; D000273:Adipose Tissue / Q000201:enzymology; D000818:Animals; D004789:Enzyme Activation; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D006168:Guinea Pigs; D006596:Hexosaminidases / Q000378:metabolism; D008071:Lipoprotein Lipase / Q000378:metabolism*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology; D011233:Precipitin Tests; D013329:Structure-Activity Relationship; D013997:Time Factors
D006023:Glycoproteins; D017485:1-Deoxynojirimycin; D008071:Lipoprotein Lipase; D006596:Hexosaminidases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D005944:Glucosamine