Biochemical and biophysical properties of a highly active recombinant arginase from Leishmania (Leishmania) amazonensis and subcellular localization of native enzyme

Show simple item record

dc.contributor.author Silva, Edson Roberto da
dc.contributor.author Laranjeira da Silva, Maria Fernanda
dc.contributor.author Fischer, Hannes
dc.contributor.author Mortara, Renato A. [UNIFESP]
dc.contributor.author Mayer, Mario Gustavo
dc.contributor.author Framesqui, Karine
dc.contributor.author Silber, Ariel Mariano
dc.contributor.author Floeter-Winter, Lucile Maria
dc.date.accessioned 2016-01-24T13:51:26Z
dc.date.available 2016-01-24T13:51:26Z
dc.date.issued 2008-06-01
dc.identifier http://dx.doi.org/10.1016/j.molbiopara.2008.02.011
dc.identifier.citation Molecular and Biochemical Parasitology. Amsterdam: Elsevier B.V., v. 159, n. 2, p. 104-111, 2008.
dc.identifier.issn 0166-6851
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/30704
dc.description.abstract Arginase (L-arginine amidinohydrolase, E.C. 3.5.3.1) is a metalloenzyme that catalyses the hydrolysis of L-arginine to L-ornithine and urea. in Leishmania spp., the biological role of the enzyme may be involved in modulating NO production upon macrophage infection. Previously, we cloned and characterized the arginase gene from Leishmania (Leishmania) amazonensis. in the present work, we successfully expressed the recombinant enzyme in E. coli and performed biochemical and biophysical characterization of both the native and recombinant enzymes. We obtained K-M and V-max. values of 23.9(+/- 0.96) mM and 192.3 mu mol/min mg protein (+/- 14.3), respectively, for the native enzyme. for the recombinant counterpart, K-M was 21.5(+/- 0.90) mM and V-max was 144.9(+/- 8.9) mu mol/min mg. Antibody against the recombinant protein confirmed a glycosomal cellular localization of the enzyme in promastigotes. Data from light scattering and small angle X-ray scattering showed that a trimeric state is the active form of the protein. We determined empirically that a manganese wash at room temperature is the best condition to purify active enzyme. the interaction of the recombinant protein with the immobilized nickel also allowed us to confirm the structural disposition of histidine at positions 3 and 324. the determined structural parameters provide substantial data to facilitate the search for selective inhibitors of parasitic sources of arginase, which could subsequently point to a candidate for leishmaniasis therapy. (c) 2008 Elsevier B.V. All rights reserved. en
dc.format.extent 104-111
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof Molecular and Biochemical Parasitology
dc.rights Acesso restrito
dc.subject protein structure en
dc.subject manganese activation en
dc.subject immunolocalization en
dc.subject drug design en
dc.title Biochemical and biophysical properties of a highly active recombinant arginase from Leishmania (Leishmania) amazonensis and subcellular localization of native enzyme en
dc.type Artigo
dc.rights.license http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institution Universidade de São Paulo (USP)
dc.contributor.institution Univ Luterana Brasil
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Inst Butantan Serv Genet
dc.description.affiliation Univ São Paulo, Dept Fisiol IB, Inst Biociencias, BR-05508900 São Paulo, Brazil
dc.description.affiliation Univ Luterana Brasil, Ctr Univ Luterano Palmas, CEULP ULBRA Palmas, São Paulo, TO, Brazil
dc.description.affiliation Univ São Paulo, Inst Fis, BR-01498 São Paulo, Brazil
dc.description.affiliation Universidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, UNIFESP, São Paulo, Brazil
dc.description.affiliation Inst Butantan Serv Genet, São Paulo, Brazil
dc.description.affiliation Univ São Paulo, Inst Ciencias Biomed, BR-05508 São Paulo, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, UNIFESP, São Paulo, Brazil
dc.identifier.doi 10.1016/j.molbiopara.2008.02.011
dc.description.source Web of Science
dc.identifier.wos WOS:000256198000004



File

File Size Format View

There are no files associated with this item.

This item appears in the following Collection(s)

Show simple item record

Search


Browse

Statistics

My Account