Cytochemical localization of ATP diphosphohydrolase from Leishmania (Viannia) braziliensis promastigotes and identification of an antigenic and catalytically active isoform.
An ATP diphosphohydrolase (EC 3.6.1.5) activity was identified in a Leishmania (Viannia) braziliensis promastigotes 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 from L. braziliensis NDPase (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 from L. braziliensis NDPase is a target for the human immune response.
Rezende-Soares
F A
FA
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil.
Carvalho-Campos
C
C
Marques
M J
MJ
Porcino
G N
GN
Giarola
N L L
NL
Costa
B L S
BL
Taunay-Rodrigues
A
A
Faria-Pinto
P
P
Souza
M A
MA
Diniz
V A
VA
Corte-Real
S
S
Juliano
M A
MA
Juliano
L
L
Vasconcelos
E G
EG
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D001081:Apyrase / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D006801:Humans; D047468:Immunoprecipitation; D007527:Isoenzymes; D007892:Leishmania braziliensis / Q000201:enzymology* / Q000276:immunology / Q000648:ultrastructure; D016773:Leishmaniasis, Cutaneous / Q000276:immunology / Q000469:parasitology*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data
D000913:Antibodies, Protozoan; D007527:Isoenzymes; D001081:Apyrase