Identification, molecular cloning and functional characterization of an octaprenyl pyrophosphate synthase in intra-erythrocytic stages of Plasmodium falciparum

dc.creatorTonhosolo, R
dc.creatorD'Alexandri, FL
dc.creatorGenta, FA
dc.creatorWunderlich, G
dc.creatorGozzo, FC
dc.creatorEberlin, MN
dc.creatorPeres, VJ
dc.creatorKimura, EA
dc.creatorKatzin, AM
dc.date2005
dc.date42309
dc.date2014-11-14T21:07:57Z
dc.date2015-11-26T17:17:00Z
dc.date2014-11-14T21:07:57Z
dc.date2015-11-26T17:17:00Z
dc.date.accessioned2018-03-29T00:05:09Z
dc.date.available2018-03-29T00:05:09Z
dc.identifierBiochemical Journal. Portland Press Ltd, v. 392, n. 117, n. 126, 2005.
dc.identifier0264-6021
dc.identifierWOS:000233542800013
dc.identifier10.1042/BJ20050441
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/69189
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/69189
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/69189
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1282409
dc.descriptionIsoprenoids play important roles in all living organisms as components of structural cholesterol, steroid hormones in mammals, carotenoids in plants, and ubiquinones. Significant differences occur in the length of the isoprenic side chains of ubiquinone between different organisms, suggesting that different enzymes are involved in the synthesis of these side chains. Whereas in Plasmodiumfalciparum the isoprenic side chains of ubiquinone contain 7-9 isoprenic units, 10-unit side chains are found in humans. In a search for the P.falciparum enzyme responsible for the biosynthesis of isoprenic side chains attached to the benzoquinone ring of ubiquinones, we cloned and expressed a putative polyprenyl synthase. Polyclonal antibodies raised against the corresponding recombinant protein confirmed the presence of the native protein in trophozoite and schizont stages of P.falciparum. The recombinant protein, as well as P.falciparum extracts, showed an octaprenyl pyrophosphate synthase activity, with the formation of a polyisoprenoid with eight isoprenic units, as detected by reverse-phase HPLC and reverse-phase TLC, and confirmed by electrospray ionization and tandem MS analysis. The recombinant and native versions of the enzyme had similar Michaelis constants with the substrates isopentenyl pyrophosphate and farnesyl pyrophosphate. The recombinant enzyme could be competitively inhibited in the presence of the terpene nerolidol. This is the first report that directly demonstrates an octaprenyl pyrophosphate synthase activity in parasitic protozoa. Given the rather low similarity of the P. falciparum enzyme to its human counterpart, decaprenyl pyrophosphate synthase, we suggest that the identified enzyme and its recombinant version could be exploited in the screening of novel drugs.
dc.description392
dc.description1
dc.description117
dc.description126
dc.languageen
dc.publisherPortland Press Ltd
dc.publisherLondon
dc.publisherInglaterra
dc.relationBiochemical Journal
dc.relationBiochem. J.
dc.rightsfechado
dc.sourceWeb of Science
dc.subjectantimalarial
dc.subjectisoprenoid
dc.subjectmalaria
dc.subjectPlasmodium falciparum
dc.subjectprenyltransferase
dc.subjectterpene
dc.subjectFarnesyl-diphosphate Synthase
dc.subjectSite-directed Mutagenesis
dc.subjectHuman Malaria Parasites
dc.subjectIsoprenoid Biosynthesis
dc.subjectConserved Aspartate
dc.subjectMevalonate Pathway
dc.subjectMass-spectrometry
dc.subjectCrystal-structure
dc.subjectEscherichia-coli
dc.subjectSide-chain
dc.titleIdentification, molecular cloning and functional characterization of an octaprenyl pyrophosphate synthase in intra-erythrocytic stages of Plasmodium falciparum
dc.typeArtículos de revistas


Este ítem pertenece a la siguiente institución