title: Cloning and characterization of two cDNAs encoding sulfatases in the Roman snail, Helix pomatia.
abstract: The sulfatase from the snail Heli pomatia is widely used for analytical applications. We have investigated the content of sulfatases in H. pomatia, using a biochemical and a molecular approach. A 112-kDa protein from the intestinal juice of H. pomatia comigrated with sulfatase activity when chromatographed on Sephacryl S300 and concanavalin A-Sepharose. The N-terminal amino acid sequence of the protein was similar to one of three sulfatase motifs defined by sequence alignment of known sulfatases. Degenerate primers designed from the motifs and the N-terminal amino acid sequence obtained were used to generate PCR fragments and to isolate both a full-length and a 3'-truncated cDNA encoding H. pomatia sulfatases, designated SULF1 and SULF2. SULF1 consists of 503 amino acids and shows 53-55% identity to the mammalian arylsulfatase B. The amino acid sequence deduced from the 878-bp SULF2 cDNA fragment is 55% identical with SULF1. Both SULF1 and SULF2 contain the cysteine residue conserved in the active site of many sulfatases, which is known to be posttranslationally modified into formylglycine in eukaryotic sulfatases. However, the SULF1 and SULF2 cDNAs do not code for the protein purified. This indicates the presence of at least three sulfatase genes in H. pomatia.
authors.affiliation: Department of Plant Biology, The Royal Veterinary and Agricultural University, Copenhagen, Denmark.
authors.forename: U
authors.initials: U
authors.lastname: Wittstock
authors.forename: M
authors.initials: M
authors.lastname: Fischer
authors.forename: I
authors.initials: I
authors.lastname: Svendsen
authors.forename: B A
authors.initials: BA
authors.lastname: Halkier
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006372:Helix, Snails / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D017386:Sequence Homology, Amino Acid; D013429:Sulfatases / Q000737:chemistry / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D013429:Sulfatases