title: Molecular cloning and functional characterization of a glucose transporter (CsGLUT) in Clonorchis sinensis.
abstract: A complementary DNA (cDNA) encoding a glucose transporter of Clonorchis sinensis (CsGLUT) was isolated from the adult C. sinensis cDNA library. The open reading frame of CsGLUT cDNA consists of 1653 base pairs that encode a 550-amino acid residue protein. Hydropathy analysis suggested that CsGLUT possess 12 putative membrane-spanning domains. The Northern blot analysis result using poly(A)(+)RNA showed a strong band at ~2.1 kb for CsGLUT. When expressed in Xenopus oocytes, CsGLUT mediated the transport of radiolabeled deoxy-D-glucose in a time-dependent but sodium-independent manner. Concentration-dependency results showed saturable kinetics and followed the Michaelis-Menten equation. Nonlinear regression analyses yielded a Km value of 588.5 ± 53.0 μM and a Vmax value of 1500.0 ± 67.5 pmol/oocyte/30 min for [1,2-(3)H]2-deoxy-D-glucose. No trans-uptakes of bile acid (taurocholic acid), amino acids (tryptophan and arginine), or p-aminohippuric acid were observed. CsGLUT-mediated transport of deoxyglucose was significantly and concentration-dependently inhibited by radio-unlabeled deoxyglucose and D-glucose. 3-O-Methylglucose at 10 and 100 μM inhibited deoxyglucose uptake by ~50 % without concentration dependence. No inhibitory effects by galactose, mannose, and fructose were observed. This work may contribute to the molecular biological study of carbohydrate metabolism and new drug development of C. sinensis.
authors.affiliation: Department of Parasitology and Tropical Medicine, Inha University School of Medicine, Incheon, 400-712, Republic of Korea.
authors.forename: Seong Kyu
authors.initials: SK
authors.lastname: Ahn
authors.affiliation: Department of Parasitology and Tropical Medicine, Inha University School of Medicine, Incheon, 400-712, Republic of Korea.
authors.forename: Pyo Yun
authors.initials: PY
authors.lastname: Cho
authors.affiliation: Department of Parasitology and Tropical Medicine, and Institute of Health Science, Gyeongsang National University School of Medicine, Jinju, 660-751, Republic of Korea.
authors.forename: Byoung-Kuk
authors.initials: BK
authors.lastname: Na
authors.affiliation: Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul, 156-756, Republic of Korea.
authors.forename: Sung-Jong
authors.initials: SJ
authors.lastname: Hong
authors.affiliation: Department of Parasitology and Catholic Institute of Parasitic Diseases, College of Medicine, Catholic University of Korea, Seoul, 137-701, Republic of Korea.
authors.forename: Ho-Woo
authors.initials: HW
authors.lastname: Nam
authors.affiliation: Department of Parasitology and Tropical Medicine, and Institute of Health Science, Gyeongsang National University School of Medicine, Jinju, 660-751, Republic of Korea.
authors.forename: Woon-Mok
authors.initials: WM
authors.lastname: Sohn
authors.affiliation: Department of Biology, Brandon University, 270-18th Street, Brandon, Manitoba, R7A 6A9, Canada.
authors.forename: Bernadette F
authors.initials: BF
authors.lastname: Ardelli
authors.affiliation: Department of Parasitology and Tropical Medicine, Inha University School of Medicine, Incheon, 400-712, Republic of Korea.
authors.forename: Yun-Kyu
authors.initials: YK
authors.lastname: Park
authors.affiliation: Department of Parasitology and Tropical Medicine, Inha University School of Medicine, Incheon, 400-712, Republic of Korea.
authors.forename: Tong-Soo
authors.initials: TS
authors.lastname: Kim
authors.affiliation: Department of Parasitology and Tropical Medicine, Inha University School of Medicine, Incheon, 400-712, Republic of Korea. shcha@inha.ac.kr.|Department of Biology, Brandon University, 270-18th Street, Brandon, Manitoba, R7A 6A9, Canada. shcha@inha.ac.kr.
authors.forename: Seok Ho
authors.initials: SH
authors.lastname: Cha
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003847:Deoxyglucose / Q000378:metabolism*; D020224:Expressed Sequence Tags; D051246:Glucose Transport Proteins, Facilitative / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D007700:Kinetics; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D010802:Phylogeny; D011061:Poly A / Q000235:genetics; D018075:RNA, Complementary / Q000378:metabolism; D018510:RNA, Helminth / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014982:Xenopus laevis
chemical_list: D018076:DNA, Complementary; D051246:Glucose Transport Proteins, Facilitative; D018075:RNA, Complementary; D018510:RNA, Helminth; D011061:Poly A; D003847:Deoxyglucose
keywords: Clonorchis sinensis; Glucose transporter; Glucose uptake; Xenopus oocyst