F A C U L T Y P R O F I L E
MANCIA, FILIPPO, Ph.D.
Structural biology of integral membrane proteins
Office: Russ Berrie Pavilion| 5th floor | Room 528
We are interested in the structure and function of membrane proteins. Proteins that reside within the plasma membrane are responsible for how a cell detects and responds to extra-cellular stimuli, of biological, chemical and physical nature. High-resolution snapshots of such molecules offer invaluable insight into the function of a given protein, and provide the framework to test structure-based mechanistic hypotheses. We primarily use X-ray diffraction of protein crystals to obtain structural information to atomic detail, while we employ multiple biophysical and biochemical techniques, as well as assays based on cells and on protein reconstituted in artificial bilayers, for functional studies. Our efforts are currently devoted to three main areas of research:
1. Insect Olfactory Receptors: Olfaction is the chief sensory modality used by many animals to find food and mates, and to avoid danger. Across all species, the first event in perceiving an odor is the binding of the odorant chemical to specific olfactory receptors (ORs). Insects, like mammals, produce a large array of ORs, and only express a single OR type in each olfactory sensory neuron. However, insects are unique in that they also express a second and invariant receptor in the vast majority of sensory neurons. Whereas variant ORs have diverged to allow species to adapt to their individual ecological niches, the second receptor, termed OR83b in Drosophila, is highly conserved and plays a key role in odor recognition. There is compelling evidence that insect variant ORs may function in complex with OR83b receptors as heteromeric ion channels. We propose to unveil the atomic resolution structure of insect ORs, and to understand how they function. The results of this research could have profound implications for human health, as it could provide the framework for the design of a strategy to halt insect borne diseases.
2. Cellular Uptake of Micronutrients: We are interested in understanding at a molecular level how essential micronutrients such as Vitamin A traverse the cell membrane. Many biological processes including vision, normal cell differentiation and proliferation, reproduction, embryonic development and immunity are crucially dependent on an adequate supply of Vitamin for proper function. Indeed, both deficiency and excess of vitamin A are associated with a wide spectrum of severe pathological phenotypes such as mental retardation, congenital heart defects and growth retardation. Vitamin A deficiency affects over 14 million children in the developing world. An understanding at a molecular level of the mechanism of Vitamin A uptake is critically dependent on high-resolution structural information, thus far lacking.
3. Structural Genomics of Membrane Proteins: We interact closely with the NIH-funded New York Consortium of Membrane Protein Structure (NYCOMPS) center located at the close-by NY Structural Biology Center, and have contributed to the design, implementation, optimization and running of a high-throughput platform for expression screening and purification of membrane proteins. With careful and rapid pre-selection of targets with increased probability of crystallization, we have been able to increase the success rate in forming crystals of membrane proteins from 0.1 to over 5%. The Mancia lab is an active participant of NYCOMPS, and receives pre-screened membrane protein clones from the Center on a regular basis.
Vasileios I. Petrou, Carmen M. Herrera, Kathryn M. Schultz, Oliver B. Clarke, Jérémie Vendome, David Tomasek, Surajit Banerjee, Kanagalaghatta R. Rajashankar, Meagan Belcher Dufrisne, Brian Kloss, Edda Kloppmann, Burkhard Rost, Candice S. Klug, M. Stephen Trent, Lawrence Shapiro, Filippo Mancia, (2016). Structures of aminoarabinose transferase ArnT suggest a molecular basis for lipid A glycosylation. Science, Vol. 351, Issue 6273, pp. 608-612.
Ardiccioni, C., Clarke, O.B., Tomasek, D., Issa, H.A., von Alpen, D.C., Pond, H.L., Banerjee, S., Rajashankar, K.R., Liu, Q., Guan, Z., Li, C., Kloss, B., Bruni, R., Kloppmann, E., Rost, B., Manzini, M.C., Shapiro, L. and Mancia, F. (2016). Structure of the polyisoprenyl-phosphate glycosyltransferase GtrB and insights into the mechanism of catalysis. Nat. Commun., 7:10175. doi: 10.1038/ncomms10175.
Clarke, O.B., Tomasek, D., Jorge, C.D., Belcher Dufrisne, M., Kim, M., Banerjee, S., Rajashankar, K.R., Shapiro, L., Hendrickson, W.A., Santos, H. and Mancia, F. (2015). Structural basis for phosphatidylinositol-phosphate biosynthesis. Nat. Commun., 6:8505. doi: 10.1038/ncomms9505.
Sciara, G., Clarke, O.B., Tomasek, D., Kloss, B., Tabuso, S., Byfield, R., Cohn, R., Banerjee, S., Rajashankar, K.R., Slavkovic, V., Graziano, J.H., Shapiro, L. and Mancia, F. (2014) Structural basis for catalysis in a CDP-alcohol phosphotransferase. Nat. Commun., 5:4068. doi: 10.1038/ncomms5068.
Sciara, G. and Mancia, F. (2012). Highlights from recently determined structures of membrane proteins: a focus on channels and transporters. Curr. Opin. Struct. Biol., 22, 476-481.
Liu, Q., Dahmane, T., Zhang, Z., Assur, Z., Brasch, J, Shapiro, L., Mancia, F. and Hendrickson, W.A. (2012) Structures from Anomalous Diffraction of Native Biological Macromolecules. Science, 336, 1033-1037.
Assur, Z., Hendrickson, W.A. and Mancia, F. (2011). Tools for Co-expressing Multiple Proteins in Mammalian Cells. Methods in Molecular Biology, 801, 173-187.
Mancia, F. and Love, J. (2011). High throughput platforms for structural genomics of integral membrane proteins. Curr. Opin. Struct. Biol., 21, 517-522.
Torres-Arancivia, C., Ross, C.M., Chavez, J., Assur, Z., Dolios, G., Mancia, F. and Ubarretxena-Belandia, I. (2010). Identification of an archaeal presenilin-like intramembrane protease. PLoS ONE, 5, e13072.
Mancia, F. and Love, J. (2010). High-throughput expression and purification of membrane proteins. J. Struct. Biol., 172, 85-93.
Love, J., Mancia, F., Shapiro, L., Punta, M., Rost, B., Girvin, M., Wang, D.N., Zhou, M., Hunt, J.F., Szyperski, T., Gouaux, E., MacKinnon, R., McDermott, A., Honig, B., Inouye, M., Montelione, G., Hendrickson, W.A. (2010). J Struct Funct Genomics, 11, 191-9.
Mancia, F., Assur, Z., Herman, A.G., Siegel, R. and Hendrickson, W.A. (2008). Asymmetry and Ligand Sensitivity in Dimeric Associations of the Serotonin 5HT2c Receptor. EMBO Rep. 9, 363-9.
Mancia, F and Hendrickson, W.A. (2007). Expression of recombinant G-protein coupled receptors for structural biology. Mol Biosyst. 3, 723-34.
Assur, Z., Schieren, I., Hendrickson, W.A. and Mancia, F. (2007). Two-color selection for amplified co-production of proteins in mammalian cells. Protein Expression and Purification, 55, 319-24.
Mancia, F., Brenner-Morton, S., Siegel, R., Assur, Z., Sun, Y., Schieren, I., Mendelsohn, M., Axel, R. and Hendrickson, W.A. (2007). Production and characterization of monoclonal antibodies sensitive to conformation in the 5HT2c serotonin receptor. Proc Natl Acad Sci U S A.104, 4303-4308.
Mancia, F., Patel, S.D., Rajala, M.W., Scherer, P.E., Nemes, A., Schieren, I., Hendrickson, W.A., and Shapiro, L. (2004). Optimization of protein production in mammalian cells with a coexpressed fluorescent marker. Structure 12, 1355-1360.