Ž . Comparative Biochemistry and Physiology Part B 130 2001 513519 Localization and characterization of acharan sulfate in the body of the giant African snail Achatina fulica Jia Jeong a , Toshihiko Toida b , Yuki Muneta b , Ichiro Kosiishi b , Toshio Imanari b , Robert J. Linhardt c , Hyung Seok Choi a , Song Ji Wu a , Yeong Shik Kim ,a a Natural Products Research Institute, Seoul National Uni  ersity, Seoul 110-460, South Korea b Faculty of Pharmaceutical Sciences, Chiba Uni  ersity, Chiba 263, Japan c Department of Chemistry, Medicinal and Natural Products Chemistry, and Chemical and Biochemical Engineering, Di  ision of Medicinal and Natural Products Chemistry, Uni  ersity of Iowa, Iowa City, IA 52242, USA Received 16 May 2001; received in revised form 17 August 2001; accepted 22 August 2001 Abstract Ž . . Ž Acharan sulfate is a glycosaminoglycan GAG , having the structure 4 -2-acetamido-2-deoxy--D-glucopyranose 1 . Ž 4 -2-sulfo--L-idopyranosyluronic acid 1 , isolated from the body of the giant African snail Achatina fulica. This GAG represents 3  5% of the dry weight of this snail’s soft body tissues. Frozen sections and polyester wax sections of Ž . the snail’s body were stained by Alcian blue-periodic acid-Schiff’s reagent PAS to localize acharan sulfate. Alcian blue staining indicated that GAG was mainly secreted into the outer surface of the body from internal granules. A highly mucous material was collected and treated and the acharan sulfate was recovered by ethanol and cetyl pyridinium chloride precipitation. Crude acharan sulfate was purified by DEAE-Sephacel ion-exchange chromatography. De- Ž . polymerization of intact mucus and purified acharan sulfate fractions by heparin lyase II heparitinase I from Fla obacterium heparinum produced an unsaturated disaccharide as a major product, establishing the repeating unit of acharan sulfate. These results demonstrate that mucus in the granule and secreted to the outside of the body is composed entirely of acharan sulfate.  2001 Elsevier Science Inc. All rights reserved. Keywords: Achatina fulica; Snail; Acharan sulfate; Localization; Section analysis; Mucus; Granules; Heparinase 1. Introduction Proteoglycans are complex macromolecules consisting of a core protein and one or more covalently attached glycosaminoglycan chains Ž . Hardingham and Fosang, 1992 . The biological functions of proteoglycans primarily result from  Corresponding author. Tel.: 82-2-740-8929; fax: 82-2- 765-4768. Ž . E-mail address: kims@plaza.snu.ac.kr Y.S. Kim . the structurally dominant glycosaminoglycan Ž . GAG chains emanating from the protein core of the molecule. A large number of animal species contain GAGs and mollusks are a particularly rich source of these polysaccharides. GAGs are usually found in the extracellular matrix of verte- brate and invertebrate tissues. A structural inves- tigation revealed that GAGs in invertebrate species often contain unusual variations of sulfate Ž distribution and uronic acids Chatziioannidis et al., 1999; Chavante et al., 2000; Pejler et al., 1096-495901$ - see front matter  2001 Elsevier Science Inc. All rights reserved. Ž . PII: S 1 0 9 6 - 4 9 5 9 01 00468-7