The focus of my research is to understand the molecular basis of MAP kinase regulation by its interaction with kinases, via scaffolding proteins, and phosphatases. We combine structural data from X-ray crystallography and NMR spectroscopy with biophysical data from ITC, CD spectroscopy and kinetics to understand how MAPK activity is controlled in vitro and in vivo. We are also investigating the structures and functions of bacterial proteins that play central roles in antibiotic resistance.

Biography

Professor Page received dual B.S. degrees with honors in Biochemistry and Applied Mathematics from the University of Arizona in 1993 and completed her graduate work in Chemistry with Dr. C. E. Schutt as an NSF graduate and Harold W. Dodds fellow at Princeton University (PhD, 2000). From 2000 - 2003, she was a NIH NRSA post-doctoral fellow at The Scripps Research Institute with Dr. Ray Stevens, after which she joined the Joint Center for Structural Genomics with Dr. Ian Wilson to become the core leader of high-throughput crystallization group. In 2005, she joined the Molecular Biology, Cell Biology and Biochemistry department at Brown University as an assistant professor. The focus of her research is to elucidate the structures and dynamics of the transient macromolecular complexes that drive signaling in eukaryotic and prokaryotic cells.

Institutions

Princeton University

Research Description

The focus of my research is to elucidate the structures and dynamics of the transient macromolecular complexes that drive signaling in eukaryotic and prokaryotic cells. Our primary tool is X-ray crystallography, which we complement with other biophysical and biochemical techniques including NMR spectroscopy, isothermal titration calorimetry, CD spectroscopy, kinetic activity assay, among others, which we use to understand how proteins direct cellular signaling at a molecular level.

The central project of the laboratory is to understand the molecular basis of MAP kinase regulation by its interaction with kinases, via scaffolding proteins, and MAPK phosphatases. Tyrosine phosphorylation is a key mechanism for the regulation of an extensive set of physiological processes. The KIM phosphatases, including hematopoietic tyrosine phosphatase (HePTP) and striatal enriched phosphatase (STEP), bind MAP kinases through a unique targeting mechanism which results in the reciprocal regulation of one another's biological activities. We are interested in understanding the molecular basis for the reciprocal regulation of the KIM phosphatase:MAP kinase interactions to determine how these interactions are modulated and directed by multiple kinase signaling pathways in the cell and how they can be regulated by developing KIM-phosphatase specific inhibitors. We are also investigating how MAPK are regulated by their interactions with scaffolding proteins in order to facilitate phosphorylation by their upstream kinases.

A second project in the laboratory is the investigation of bacterial proteins that play a central role in biofilm formation and antibiotic resistance. In recent years, genes important for . E coli biofilm formation and propagation have been identified using DNA-array experiments, proteome analysis, and classical knockout studies. Identified genes code for proteins involved in bacterial motility, quorum sensing, and the induction of polysaccharide synthesis. Unfortunately, the specific biochemical functions of many of these genes are unknown. Because protein function is more highly correlated with structure than sequence, we are using X-ray crystallography and NMR spectroscopy to determine the atomic resolution structures of these proteins and protein complexes to gain insights into their individual and coordinated functions in antibiotic resistance and biofilm formation. As an example, we recently used X-ray crystallography, biochemistry and genetic to show that the protein most upregulated in the bacterial persister cells (these cells are a phenotypically distinct yet genetically identical subset of cells that exhibit high tolerance to antibiotics), MqsR, along with MqsA, are the founding members of an entirely novel toxin:antitoxin system in E. coli; this work has provided a basis for developing novel antibacterial therapies that target TA pairs.

Awards

• 2008-2011 American Cancer Society Research Scholar
  
• 2005 Rhode Island Foundation Medical Research Grant
  
• 2005 Richard B. Salomon Faculty Research Award
  
• 2000-2003 Ruth L. Kirschstein National Research Service Award (NRSA), NIH
  
• 1997-1998 Harold W. Dodds Honorific Graduate Fellowship
• 1994-1997 National Science Foundation Graduate Fellowship
• 1993-1994 Phi Beta Kappa
  
• 1992-1993 Barry M. Goldwater Undergraduate Research Scholar
• 1988-1992 Regent's Academic Achievement Scholarship

Affiliations

• 2007-present Executive Board Member, Protein Expression Research Group, ABRF/FASEB
  
• 2005–present Associate Editor, Protein Expression and Purification
  
• 2004-present American Crystallographic Association

Funded Research

• Structural Basis of T Cell Regulation by HePTP
  American Cancer Society (RSG-08-067-01-LIB)
  01/01/2008 - 12/31/2011
  PI: Rebecca Page
  
• Expression of proteins for structural studies and drug development
  Schering-Plough
  6/15/2008 - 12/31/2010
  PI: Rebecca Page
  
• Structural Biology and Function of Macromolecular Complexes
  Research Seed Fund Award, Brown University
  02/01/2006 - 12/31/2006
  PI: Rebecca Page
  
  

Courses Taught

• Advanced Biochemistry (Biol 1270/2270)
• Current Topics in Biochemistry and Molecular Biology (Biol2200)

Selected Publications

• Kelker, M.S., Page, R.*, Peti, W.* (2009) Crystal Structures of Protein Phosphatase-1 Bound to Nodularin –R and Tautomycin: a novel scaffold for structure based drug design of serine/threonine phosphatase inhibitors. J Mol Biol, 382, 11-21. *Contributing authors.(2009)
• Francis, D.M., Page R. (2009) Strategies to Optimize Protein Expression in E. coli. Current Protocols in Protein Chemistry (in press).(2009)
• Brown, B.L., Hadley, M., and Page, R. (2008) Heterologous high-level E. coli expression, purification and biophysical characterization of the spine-associated RapGAP (SPAR) PDZ domain. Prot Expr Purif, 62, 9-14.(2008)
• Dugan, A.S., Maginnis, M.S., Jordan, J.A., Gasparovic, M.L., Manley, K., Page, R., Williams, G., Porter, E. O'Hara, B.A., and Atwood, W.J. (2008) Human alpha-defensins inhibit BK virus infection by aggregating virions and blocking binding to host cells. J Biol Chem, 283, 31125-31132.(2008)
• Critton, D.A., Tortajada, A., Stetson, G., Peti, W., Page, R. (2008). Structural basis of substrate recognition by Hematopoietic Tyrosine Phosphatase (HePTP). Biochemistry, 47, 13336-13345. This work was recognized as a highlighted article, ACS Biochemistry and highlighted by the Faculty of 1000, Biology (http://www.f1000biology.com/article/id/1157920/evaluation.(2008)
• Lee, J.*, Page, R.*, Contreras, R.G., Palermino, J.M., Zhang, X.S., Doshi, O., Wood, T.K. and Peti, W. (2007) Structure and function of the E. coli protein YMGB : a protein critical for biofilm formation and acid resistance, J Mol Biol, 373, 11-26. *These authors contributed equally to this work; this work has been highlighted by the Faculty of 1000, Biology (http://www.f1000biology.com/article/id/1094851/evaluation).(2007)
• Page, R.*, Peti, W.*, Wilson, I. A., Stevens, R. C., & Wüthrich, K. (2005). NMR screening of mouse homologue proteins in a structural proteomics pipeline. PNAS, 102, 1901-1905. *These authors contributed equally to this work. This work has been highlighted by the Faculty of 1000, Biology (http://www.f1000biology.com/article/id/1094851/evaluation).(2005)
• Mustelin, T., Tautz, L. and Page, R. (2005). Structure of the Hematopoietic Tyrosine Phosphatase (HePTP) Catalytic Domain: Structure of a KIM phosphatase with a phosphate bound at the active site. J Mol Biol, 354, 150-163 .(2005)
• Peti, W., Johnson, M.A., Herrmann, T., Neuman, B.W., Buchmeier, M.J., Nelson, M., Joseph, J., Page, R., Stevens, R.C., Kuhn, P. & Wüthrich, K. (2005) Structural Genomics of the SARS Coronavirus: NMR Structure of the Protein nsP7. J Virology, 79, 12905-12913.(2005)
• Collins, B., Stevens, R.C., Page, R. (2005) High-Throughput Optimum Solubility Screening: Using Crystallization Results to Identify the Optimal Buffer for Protein Crystal Formation. Acta Crystallogr F61, 1035-1038.(2005)
• Page, R., Nelson, M. S., von Delft, F., Elsliger, M. A., Canaves, J. M., Brinen, L. S., Dai, X., Deacon, A. M., Floyd, R., Godzik, A., Grittini, C., Grzechnik, S. K., Jaroszewski, L., Klock, H. E., Koesema, E., Kovarik, J. S., Kreusch, A., Kuhn, P., Lesley, S. A., McMullan, D., McPhillips, T. M., Miller, M. D., Morse, A., Moy, K., Ouyang, J., Robb, A., Rodrigues, K., Schwarzenbacher, R., Spraggon, G., Stevens, R. C., van den Bedem, H., Velasquez, J., Vincent, J., Wang, X., West, B., Wolf, G., Hodgson, K. O., Wooley, J. & Wilson, I. A. (2004). Crystal structure of gamma-glutamyl phosphate reductase (TM0293) from Thermotoga maritima at 2.0 Å resolution. Proteins 54, 157-161(2004)
• Page, R & Stevens, R. C. (2004). Crystallization data mining in structural genomics: using positive and negative results to optimize protein crystallization screens. Methods, 34, 373-389.(2004)
• Page, R. Moy, K., Sims, E. C., Velasquez, J., McManus, B., Grittini, C & Stevens, R. C. (2004). Scalable high-throughput microliter expression device for Escherichia coli recombinant proteins. Biotechniques, 37, 364-370.(2004)
• Canaves, J., Page, R. & Stevens, R. C. (2004) Protein Biophysical Properties that Correlate with Crystallization Success in Thermotoga maritima: maximum clustering strategy for structural genomics. J Mol Biol, 344, 977-991.(2004)
• Peti, W., Page, R., Moy, K., Stevens, R. C. & Wüthrich, K. (2004). Miniaturization of high-throughput NMR screening for structural proteomics. J Struct Funct Genomics, 6, 209-217.(2004)
• Page, R., Grzechnik, S. K., Canaves, J. M., Spraggon, G., Kreusch, A., Kuhn, P., Stevens, R. C. & Lesley, S. A. (2003). Shotgun crystallization strategy for structural genomics: an optimized two-tiered crystallization screen against the Thermotoga maritima proteome. Acta Crystallogr D Biol Crystallogr 59, 1028-1037(2003)
• Nyman, T., Page, R., Schutt, C. E., Karlsson, R. & Lindberg, U. (2002). A cross-linked profilin-actin heterodimer interferes with elongation at the fast-growing end of F-actin. J Biol Chem 277, 15828-15833.(2002)
• Page, R., Lindberg, U. & Schutt, C. E. (1998). Domain motions in actin. J Mol Biol 280, 463-474.(1998)