Cytochemical localization of ATP diphosphohydrolase fromLeishmania (Viannia) braziliensispromastigotes and identification of an antigenic and catalytically active isoform
| Autor: | Maria A. Juliano, A. Taunay-Rodrigues, Luiz Juliano, Gabriane Nascimento Porcino, B. L. S. Costa, Maria Aparecida de Souza, Eveline Gomes Vasconcelos, Priscila Faria-Pinto, Suzana Corte-Real, Cristiane de Carvalho-Campos, F. A. Rezende-Soares, N. L. L. Giarola, Marcos José Marques, Vanessa Alvaro Diniz |
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| Rok vydání: | 2009 |
| Předmět: |
Blotting
Western Molecular Sequence Data Antibodies Protozoan Leishmaniasis Cutaneous Leishmania braziliensis Epitope Cutaneous leishmaniasis Antigen medicine Animals Humans Immunoprecipitation Amino Acid Sequence biology Apyrase Leishmania biology.organism_classification medicine.disease Molecular biology Isoenzymes Microscopy Electron Infectious Diseases Polyclonal antibodies biology.protein Animal Science and Zoology Parasitology Antibody |
| Zdroj: | Parasitology. 137:773-783 |
| ISSN: | 1469-8161 0031-1820 |
| Popis: | SUMMARYAn ATP diphosphohydrolase (EC 3.6.1.5) activity was identified in aLeishmania (Viannia) braziliensispromastigotes preparation (Lb). Ultrastructural cytochemical microscopy showed this protein on the parasite surface and also stained a possible similar protein at the mitochondrial membrane. Isolation of an active ATP diphosphohydrolase isoform from Lb was obtained by cross-immunoreactivity with polyclonal anti-potato apyrase antibodies. These antibodies, immobilized on Protein A-Sepharose, immunoprecipitated a polypeptide of approximately 48 kDa and, in lower amount, a polypeptide of approximately 43 kDa, and depleted 83% ATPase and 87% of the ADPase activities from detergent-homogenized Lb. Potato apyrase was recognized in Western blots by IgG antibody from American cutaneous leishmaniasis (ACL) patients, suggesting that the parasite and vegetable proteins share antigenic conserved epitopes. Significant IgG seropositivity in serum samples diluted 1:50 from ACL patients (n=20) for Lb (65%) and potato apyrase (90%) was observed by ELISA technique. Significant IgG antibody reactivity was also observed against synthetic peptides belonging to a conserved domain fromL. braziliensisNDPase (80% seropositivity) and its potato apyrase counterpart (50% seropositivity), in accordance with the existence of shared antigenic epitopes and demonstrating that in leishmaniasis infection the domain r82-103 fromL. braziliensisNDPase is a target for the human immune response. |
| Databáze: | OpenAIRE |
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