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J Proteomics


Title:Expression of the mevalonate pathway enzymes in the Lutzomyia longipalpis (Diptera: Psychodidae) sex pheromone gland demonstrated by an integrated proteomic approach
Author(s):Gonzalez-Caballero N; Rodriguez-Vega A; Dias-Lopes G; Valenzuela JG; Ribeiro JM; Carvalho PC; Valente RH; Brazil RP; Cuervo P;
Address:"Laboratorio de Bioquimica e Fisiologia de Insetos, IOC, FIOCRUZ, Rio de Janeiro, RJ, Brazil. Laboratorio de Pesquisa em Leishmaniose, IOC, FIOCRUZ, Rio de Janeiro, RJ, Brazil. Pos-graduacao Biologia Parasitaria, IOC, FIOCRUZ, Rio de Janeiro, RJ, Brazil. Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institutes of Health Rockville, MD, USA. Vector Biology Section, Laboratory of Malaria and Vector Research, National Institutes of Health Rockville, MD, USA. Laboratorio de Proteomica e Engenharia de Proteinas, Instituto Carlos Chagas, FIOCRUZ, Curitiba, Brazil. Laboratorio de Toxinologia, IOC, FIOCRUZ, Rio de Janeiro, RJ, Brazil. Laboratorio de Pesquisa em Leishmaniose, IOC, FIOCRUZ, Rio de Janeiro, RJ, Brazil. Electronic address: pcuervo@fiocruz.br"
Journal Title:J Proteomics
Year:2014
Volume:20131101
Issue:
Page Number:117 - 132
DOI: 10.1016/j.jprot.2013.10.028
ISSN/ISBN:1876-7737 (Electronic) 1874-3919 (Print) 1874-3919 (Linking)
Abstract:"In Latin America, Lutzomyia longipalpis is the main vector of the protozoan parasite Leishmania infantum, which is the causal agent of American Visceral Leishmaniasis. This insect uses male-produced pheromones for mate recognition. Elucidation of pheromone biogenesis or its regulation may enable molecular strategies for mating disruption and, consequently, the vector's population management. Motivated by our recent results of the transcriptomic characterization of the L. longipalpis pheromone gland, we performed a proteomic analysis of this tissue combining SDS-PAGE, and mass spectrometry followed by an integrative data analysis. Considering that annotated genome sequences of this sand fly are not available, we designed an alternative workflow searching MS/MS data against two customized databases using three search engines: Mascot, OMSSA and ProLuCID. A total of 542 proteins were confidently characterized, 445 of them using a Uniref100-insect protein database, and 97 using a transcript translated database. In addition, use of PEAKS for de novo peptide sequencing of MS/MS data confirmed ~90% identifications made with the combination of the three search engines. Our results include the identification of six of the seven enzymes of the mevalonate-pathway, plus the enzymes involved in sesquiterpenoid biosynthesis, all of which are proposed to be involved in pheromone production in L. longipalpis. BIOLOGICAL SIGNIFICANCE: L. longipalpis is the main vector of the protozoan parasite L. infantum, which is the causal agent of American Visceral Leishmaniasis. One of the control measures of such disease is focused on vector population control. As this insect uses male-produced pheromones for mate recognition, the elucidation of pheromone biogenesis or its regulating process may enable molecular strategies for mating disruption and, consequently, this vector's population management. On this regard, in this manuscript we report expression evidence, at the protein level, of several molecules potentially involved in the pheromone production of L. longipalpis. Our results include the identification of the mevalonate-pathway enzymes, plus the enzymes involved in sesquiterpenoid biosynthesis, all of which are proposed to be involved in pheromone production in L. longipalpis. In addition, considering that the annotated genome sequences of this sand fly are not yet available, we designed an alternative workflow searching MS/MS data against proteomic and transcript translated customized databases, using three search engines: Mascot, OMSSA, and ProLuCID. In addition, a de novo peptide sequencing software (PEAKS) was used to further analyze the MS/MS data. This approach made it possible to identify and annotate 542 proteins for the pheromone gland of L. longipalpis. Importantly, all annotated protein sequences and raw data are available for the research community in protein repositories that provide free access to the data"
Keywords:"Animals Diptera/*enzymology/genetics/parasitology Exocrine Glands/*enzymology Female Gene Expression Regulation, Enzymologic Insect Proteins/*biosynthesis/genetics Insect Vectors/*enzymology/genetics/parasitology Leishmania infantum Leishmaniasis, Viscera;"
Notes:"MedlineGonzalez-Caballero, Natalia Rodriguez-Vega, Andres Dias-Lopes, Geovane Valenzuela, Jesus G Ribeiro, Jose M C Carvalho, Paulo Costa Valente, Richard H Brazil, Reginaldo P Cuervo, Patricia eng ZIA AI000810-16/Intramural NIH HHS/ Research Support, N.I.H., Intramural Research Support, Non-U.S. Gov't Netherlands 2013/11/05 J Proteomics. 2014 Jan 16; 96:117-32. doi: 10.1016/j.jprot.2013.10.028. Epub 2013 Nov 1"

 
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