title: Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae).
abstract: Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. On the other hand, the hemorrhagic vascular lesions observed are attributed to the proteolytic activity of the toxin. Thus, N-glycan sulfation may constitute a defense mechanism against predators.
authors.affiliation: Department of Carbohydrates, Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana, Cuba.
authors.forename: Gleysin
authors.initials: G
authors.lastname: Cabrera
authors.affiliation: Centro de Biofísica y Bioquímica.
authors.forename: Víctor
authors.initials: V
authors.lastname: Salazar
authors.affiliation: School of Biological Sciences, Universidad de Concepción, Víctor Lamas 1290, PO Box 160C, Concepción, Chile.
authors.forename: Raquel
authors.initials: R
authors.lastname: Montesino
authors.affiliation: Department of Proteomics, Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana, Cuba.
authors.forename: Yanet
authors.initials: Y
authors.lastname: Támbara
authors.affiliation: Department of Chemistry, University of Oxford, Chemistry Research Laboratory, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Weston B
authors.initials: WB
authors.lastname: Struwe
authors.affiliation: Proteomics Unit, Center of Structural Biology.
authors.forename: Evelyn
authors.initials: E
authors.lastname: Leon
authors.affiliation: Glycobiology Institute, Department of Biochemistry, Oxford University, South Parks Road, Oxford OX1 3QU, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Harvey
authors.affiliation: Luxembourg Clinical Proteomics Center, 1A-B, rue Thomas Edison, L-1445 Strassen, Luxembourg.
authors.forename: Antoine
authors.initials: A
authors.lastname: Lesur
authors.affiliation: Proteomics Unit, Center of Structural Biology.
authors.forename: Mónica
authors.initials: M
authors.lastname: Rincón
authors.affiliation: Luxembourg Clinical Proteomics Center, 1A-B, rue Thomas Edison, L-1445 Strassen, Luxembourg.
authors.forename: Bruno
authors.initials: B
authors.lastname: Domon
authors.affiliation: Proteomics Unit, Center of Structural Biology.
authors.forename: Milagros
authors.initials: M
authors.lastname: Méndez
authors.affiliation: Unidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo, Mataojo 2020, Montevideo, Uruguay.
authors.forename: Madelón
authors.initials: M
authors.lastname: Portela
authors.affiliation: Department of Carbohydrates, Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana, Cuba.
authors.forename: Annia
authors.initials: A
authors.lastname: González-Hernández
authors.affiliation: Department of Carbohydrates, Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana, Cuba.
authors.forename: Ada
authors.initials: A
authors.lastname: Triguero
authors.affiliation: Unidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo and IIBCE, Mataojo 2020, Montevideo, Uruguay.
authors.forename: Rosario
authors.initials: R
authors.lastname: Durán
authors.affiliation: Unit for Invertebrate Toxins, Venezuelan Institute for Scientific Research (IVIC), PO Box 20632, Caracas 1020A, Venezuela.
authors.forename: Ulf
authors.initials: U
authors.lastname: Lundberg
authors.affiliation: Proteomics Unit, Center of Structural Biology.
authors.forename: Eva
authors.initials: E
authors.lastname: Vonasek
authors.affiliation: Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana, Cuba luis.javier@cigb.edu.cu.
authors.forename: Luis Javier
authors.initials: LJ
authors.lastname: González
mesh_terms: D000818:Animals; D006031:Glycosylation; D009036:Moths / Q000201:enzymology; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000737:chemistry* / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D057057:Serine Proteases / Q000737:chemistry* / Q000378:metabolism; D013431:Sulfates / Q000737:chemistry / Q000378:metabolism
chemical_list: D011134:Polysaccharides; D013431:Sulfates; D057057:Serine Proteases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
keywords: Hylesia metabus; insects; lepidopterism; serine protease; sulfated N-glycans