Mario Tyago Murakami
|Since 08/2008Principal Investigator, LNBio-CNPEMMechanistic & Structural Biology GroupCoordinator: Macromolecular Crystallization Facility (LNBio) and MX2 Beamline (LNLS)
Phone: +55 19 3512.1267
Our research focuses on the elucidation of the molecular and mechanistic basis of biological phenomena related to intracellular trafficking in eukaryotic cells, plant cell-wall breakdown and trypanosomatid-related diseases. For this purpose, we employ a multi-disciplinary approach which combines high-resolution biophysical methods (X-ray crystallography and NMR), recombinant DNA technologies (cloning, mutagenesis and yeast two hybrid), enzymology, calorimetry and cell-based assays.
Intracellular trafficking mediated by actin-based molecular motors
Unconventional myosins are actin-based molecular motors ubiquitous in eukaryotic cells, which participate in a multitude of intracellular processes including cell division, cell movement, intracellular transport and signal transduction. Mutations or defects in myosin genes are related to a variety of human diseases such as Griscelli syndrome that is characterized by hypopigmentation and neurological disorders and some types of cancer promoting proliferation, invasion and motility. The goal of this research line is shed light on the sophisticated molecular mechanisms involved in the signaling, regulation and selectivity of these essential molecular motors for eukaryotes.
Structure-function-stability relationships in multi-domain glycosyl hydrolases
Glycoside Hydrolases (GHs) catalyze the hydrolysis of glycosidic bonds between carbohydrate and/or aglycone moieties. These enzymes are found in all living organisms playing key roles in different biological activities such as plant cell-wall construction and breakdown, pathogenesis mechanisms, normal cellular function and anti-bacterial defense strategies. Besides their importance for multiple physiological events, GHs have also been extensively employed in a variety of biotechnological processes such as biobleaching of paper and pulp, food processing, organic synthesis and recently in the reduction of lignocellulosic biomass into fermentable sugars for biofuels production. Our group has been working on the structural and functional aspects of industrially-relevant GHs aiming at elucidating molecular mechanisms involved in specificity, operation mode, thermal stability and regulation. We are also interested in determining how native and “foreign” non-catalytic modules can affect enzyme stability and function. Finally, we expect to combine/correlate all these functional and structural findings at molecular level to aid the rational (re)design of enzymes for the different biotechnological purposes.
Functional and Structural Studies of Protein Kinases involved in neglected diseases
Protein kinases (PKs) play key roles in the cell cycle of trypanosomatids and recent studies demonstrated their importance for parasite growth and survival. In this collaborative project with Profs. Jorg Kobarg and Artur Cordeiro, we are focused on PKs from Leishmania braziliensis and Trypanosoma cruzi, species endemic to Latin America which causes mucocutaneous leishmaniasis and Chagas diseases, respectively. Our targets include PKs belonging to the NEK (NIMA related kinases) family and to the CMGC, AGC and other PKs groups. Literature indicates them as promising targets for drug discovery and development against human pathogenic trypanosomatids. The mission of this research is to contribute for a better understanding of the molecular and biological function of PKs from trypanosomatids of medical interest and further in the discovery and development of new drugs for the treatment of trypanosomiasis. For this purpose, we use a target-based approach which combines protein crystallography, HTS, virtual screening, ITC and cell-based assays for target validation, hits identification and confirmation.
SELECTED PUBLICATIONS (2012)
Santos, C.R., Polo, C.C., Correa, J.M., Simao, R.G.G., Seixas, F.A.V. , Murakami, M.T. Accessory domain changes accessibility and molecular topography of the catalytic interface in monomeric GH39 beta-xylosidases. Acta Crystallographica D, Biological Crystallography, 2012. (In press) (IF – 12.6)
Santos, C. R., Paiva, J. H., Sforca, M.L., Neves, J.L., Navarro, R.Z., Cota-Silva, J., Akao, P. K., Hoffmam, Z. B., Meza, A.N., Smetana, J., Nogueira, M.L.C., Polikarpov, I., Xavier-Neto, J., Squina, F.M., Ward, R. J., Ruller, R., Zeri, A. C., Murakami, M.T. Dissecting structure-function-stability relationships of a thermostable GH5-CBM3 cellulase from Bacillus subtilis 168. Biochemical Journal, v.441, p.95-104, 2012. (IF – 4.9)
Santos, C.R., Paiva, J.H., Meza, A.N., Cota, J., Alvarez, T.M., Ruller, R., Prade, R.A., Squina, F.M., Murakami, M.T. Molecular insights into substrate specificity and thermal stability of a bacterial GH5-CBM27 endo-1,4-b-d-mannanase. Journal of Structural Biology, v. 177, p. 469-476, 2012. (IF – 3.4)
Giuseppe, P.O., Martins, N.H., Navarro, A.M., Santos, C.R., Pereira, H.M., Murakami, M.T. Insights into phosphate cooperativity and influence of substrate modifications on binding and catalysis of hexameric purine nucleoside phosphorylases. Plos One, 2012. (In press) (IF – 4.1)
Segato, F., Damasio, A.R.L., Goncalves, T.A., Murakami, M.T., Squina, F.M., Polizeli, Maria, L.T.M., Mort, A., Prade, R.A. Two structurally discrete GH7-cellobiohydrolases compete for the same cellulosic substrate fiber. Biotechnology Biofuels, v. 5, p. 21, 2012. (IF – 6.1)
EDUCATION & TRAINING
05/2012-07/2012 Postdoctoral Research at Rutgers University, USA. Biomolecular NMR. Supervisor: Prof. Dr. Kalodimos (Babis) Charalampos.
01/2007-10/2007 Postdoctoral Research at UNESP, Brazil. Macromolecular Crystallography. Supervisor: Prof. Dr. Raghuvir Krishnaswamy Arni.
02/2003-12/2006 PhD Thesis: “Strategies of inhibition and interactions in macromolecular complexes”, UNESP, Brazil. Supervisor: Prof. Dr. Raghuvir Krishnaswamy Arni.
07/2005-02/2006 Sandwich-Doctorate at University of Hamburg, Germany. Protein Crystallography. Supervisor: Prof. Dr. Christian Betzel.
01/2000-12/2002 Undergraduate project: Structure and function of hemostatically-active proteases, UNESP, Brazil. Supervisor: Prof. Dr. Raghuvir Krishnaswamy Arni.
07/2007 – 08/2008 Collaborating Professor at the Department of Physics, IBILCE/UNESP, Brazil.