Determination of three-dimensional structure, characterization of natural substrate and cellular localization of luciferase enzyme-like (protoluciferase) of Zophobas morio

• Grant number: 12/02161-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: June 01, 2012
• End date: May 31, 2015
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal Investigator: Vadim Viviani
• Grantee: Rogilene Aparecida Prado
• Host Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil

Abstract

Currently, due to the new sequencing methods, the genome and transcriptome of hundreds of species are available in databases, and the number of sequences generated by this new technology increases every day. However, the interpretation of such sequence information does not follow the same pace. The prediction of protein function remains a challenge. Although the prediction of protein function of new sequences is based on the comparison with the sequences of proteins with previously assigned and annotated functions in databases, the specific function of thousands of proteins remains unknown. An example is the large amount of luciferase-like enzymes in many organisms. Beetle luciferases are enzymes belonging to the superfamily of AMP/ CoA ligases, enzymes that catalyze the adenylation and thioesterification of a wide variety of carboxylic substrates with CoA. Recently we cloned the first luminescent luciferase-like enzyme from the Malpighian tubules of Zophobas (Tenebrionidae) mealworm. Although this enzyme has been cloned from a non-luminescent beetle and has a low identity with beetle luciferases, it displays a weak luminescence in the presence of D-luciferin and MgATP, the substrates of the bioluminescent reaction of beetles, with a peak at ~ 613 nm. Thus, this enzyme has become a good model for studying the origin and structural evolution of beetle luciferase activity, one of the most intriguing issues about bioluminescence. Despite the kinetic characterization of the luminescent reaction of this enzyme has been carried out, the natural carboxylic substrate and the biological function of this enzyme in the Malpighian tubules are still unknown. Therefore, this project aims to identify the natural substrate of this enzyme, its cellular localization and determination of three-dimensional structure. The results will help to identify the enzymatic function of this new luciferase-like enzyme in the Malpighian tubules of beetles, helping to determine the function of other luciferase-like enzymes. (AU)