Glycogenin-2, a novel self-glucosylating protein involved in liver glycogen biosynthesis.
Glycogenin is a self-glucosylating protein involved in the initiation phase of glycogen biosynthesis. A single mammalian gene had been reported to account for glycogen biogenesis in liver and muscle, the two major repositories of glycogen. We describe the characterization of novel forms of glycogenin, designated glycogenin-2 (GN-2), encoded by a second gene that is expressed preferentially in certain tissues, including liver, heart, and pancreas. Cloning of cDNAs encoding glycogenin-2 indicated the existence of multiple species, including three liver forms (GN-2alpha, GN-2beta, and GN-2gamma) generated in part by alternative splicing. Overall, GN-2 has 40-45% identity to muscle glycogenin but is 72% identical over a 200-residue segment thought to contain the catalytic domain. GN-2 expressed in Escherichia coli or COS cells is active in self-glucosylation assays, and self-glucosylated GN-2 can be elongated by skeletal muscle glycogen synthase. Antibodies raised against GN-2 produced in E. coli recognized proteins of Mr approximately 66,000 present in extracts of rat liver and in cultured H4IIEC3 hepatoma cells. In H4IIEC3 cells, most of the GN-2 was present as a free protein but some was covalently associated with glycogen fractions and was only released by treatment with alpha-amylase. H4IIEC3 cells also expressed the muscle form of glycogenin (glycogenin-1), which was attached to a chromatographically separable glycogen fraction.
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Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana 46202-5122, USA.
Skurat
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Roach
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PJ
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D005964:Glucosyltransferases; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006031:Glycosylation; D008112:Liver Glycogen / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D051381:Rats; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D006023:Glycoproteins; D008112:Liver Glycogen; D005964:Glucosyltransferases; C109150:GYG2 protein, human