title: Leishmania (Viannia) braziliensis: insights on subcellular distribution and biochemical properties of heparin-binding proteins.
abstract: Leishmaniasis is a vector-borne disease and an important public health issue. Glycosaminoglycan ligands in Leishmania parasites are potential targets for new strategies to control this disease. We report the subcellular distribution of heparin-binding proteins (HBPs) in Leishmania (Viannia) braziliensis and specific biochemical characteristics of L. (V.) braziliensis HBPs. Promastigotes were fractionated, and flagella and membrane samples were applied to HiTrap Heparin affinity chromatography columns. Heparin-bound fractions from flagella and membrane samples were designated HBP Ff and HBP Mf, respectively. Fraction HBP Ff presented a higher concentration of HBPs relative to HBP Mf, and SDS-PAGE analyses showed 2 major protein bands in both fractions (65 and 55 kDa). The 65 kDa band showed gelatinolytic activity and was sensitive to inhibition by 1,10-phenanthroline. The localization of HBPs on the promastigote surfaces was confirmed using surface plasmon resonance (SPR) biosensor analysis by binding the parasites to a heparin-coated sensor chip; that was inhibited in a dose-dependent manner by pre-incubating the parasites with variable concentrations of heparin, thus indicating distinct heparin-binding capacities for the two fractions. In conclusion, protein fractions isolated from either the flagella or membranes of L. (V.) braziliensis promastigotes have characteristics of metallo-proteinases and are able to bind to glycosaminoglycans.
authors: Laboratório de Biologia Molecular e Doenças Endêmicas, IOC-FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, CEP 21040-360, Brasil.
authors: Luzia Monteiro
authors: LM
authors: de Castro Côrtes
authors: Mirian Claudia
authors: MC
authors: de Souza Pereira
authors: Francisco Odêncio Rodrigues
authors: FO
authors: de Oliveira
authors: Suzana
authors: S
authors: Corte-Real
authors: Franklin Souza
authors: FS
authors: da Silva
authors: Bernardo Acácio Santini
authors: BA
authors: Pereira
authors: Maria
authors: M
authors: de Fátima Madeira
authors: Marcia Terezinha Baroni
authors: MT
authors: de Moraes
authors: Reginaldo Peçanha
authors: RP
authors: Brazil
authors: Carlos Roberto
authors: CR
authors: Alves
mesh_terms: D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D002458:Cell Fractionation; D005786:Gene Expression Regulation / Q000502:physiology*; D007892:Leishmania braziliensis / Q000502:physiology* / Q000648:ultrastructure; D010447:Peptide Hydrolases / Q000378:metabolism; D021381:Protein Transport
chemical_list: D015815:Cell Adhesion Molecules; C074285:heparin-binding protein (Leishmania); D010447:Peptide Hydrolases