Title

Cloning of a mouse beta 1,3 N-acetylglucosaminyltransferase GlcNAc(beta 1,3)Gal(beta 1,4)Glc-ceramide synthase gene encoding the key regulator of lacto-series glycolipid biosynthesis

UMMS Affiliation

Eunice Kennedy Shriver Center

Date

6-1-2001

Document Type

Article

Subjects

Amino Acid Motifs; Amino Acid Sequence; Amino Sugars; Animals; B-Lymphocytes; Base Sequence; Blotting, Northern; Brain; Cell Line; Cerebellum; Chromatography, Thin Layer; Cloning, Molecular; DNA, Complementary; Glycolipids; In Situ Hybridization; Insects; Mice; Models, Chemical; Molecular Sequence Data; N-Acetylglucosaminyltransferases; Open Reading Frames; Recombinant Proteins; Spleen; Substrate Specificity; Time Factors; Tissue Distribution

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

The distinction between the different classes of glycolipids is conditioned by the action of specific core transferases. The entry point for lacto-series glycolipids is catalyzed by the beta1,3 N-acetylglucosaminyltransferase GlcNAc(beta1,3)Gal(beta1,4)Glc-ceramide (Lc3) synthase enzyme. The Lc3 synthase activity has been shown to be regulated during development, especially during brain morphogenesis. Here, we report the molecular cloning of a mouse gene encoding an Lc3 synthase enzyme. The mouse cDNA included an open reading frame of 1131 base pairs encoding a protein of 376 amino acids. The Lc3 synthase protein shared several structural motifs previously identified in the members of the beta1,3 glycosyltransferase superfamily. The Lc3 synthase enzyme efficiently utilized the lactosyl ceramide glycolipid acceptor. The identity of the reaction products of Lc3 synthase-transfected CHOP2/1 cells was confirmed by thin-layer chromatography immunostaining using antibodies TE-8 and 1B2 that recognize Lc3 and Gal(beta1,4)GlcNAc(beta1,3)Gal(beta1,4)Glc-ceramide (nLc4) structures, respectively. In addition to the initiating activity for lacto-chain synthesis, the Lc3 synthase could extend the terminal N-acetyllactosamine unit of nLc4 and also had a broad specificity for gangliosides GA1, GM1, and GD1b to generate neolacto-ganglio hybrid structures. The mouse Lc3 synthase gene was mainly expressed during embryonic development. In situ hybridization analysis revealed that that the Lc3 synthase was expressed in most tissues at embryonic day 11 with elevated expression in the developing central nervous system. Postnatally, the expression was restricted to splenic B-cells, the placenta, and cerebellar Purkinje cells where it colocalized with HNK-1 reactivity. These data support a key role for the Lc3 synthase in regulating neolacto-series glycolipid synthesis during embryonic development.

Rights and Permissions

Citation: J Biol Chem. 2001 Aug 10;276(32):30261-9. Epub 2001 May 30. Link to article on publisher's site

DOI of Published Version

10.1074/jbc.M102979200

Related Resources

Link to Article in PubMed

Journal Title

The Journal of biological chemistry 1,3-N-acetyl-beta-D-glucosaminyltransferase)

PubMed ID

11384981