Rebecca Page, Principal Investigator

Department of Molecular Biology, Cell Biology and Biochemistry

My laboratory uses X-ray crystallography to understand the molecular basis of protein function, with a particular interest in understanding how scaffolding proteins and tyrosine phosphatases regulate neuronal signaling and T-cell proliferation and how bacterial signaling proteins regulate biolfim formation.

Map Kinase Tyrosine 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), 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.

Dendritic Spines

Scaffolding proteins are multidomain proteins that organize large macromolecular assemblies in the cell. In the dendritic spine, scaffolding proteins are responsible for linking the proteins at the cell membrane, ion channels and transmembrane receptors, to the proteins within the cell, including phosphate signaling proteins and those of the actin cytoskeleton. We want to understand the detailed interactions of scaffolding proteins, both alone and as complexes with their multiple binding partners, so we can begin to elucidate how chemical signals from outside the membrane are transduced to the signaling pathways inside the cell.

Bacterial Biofilms

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 versus 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 resulting in a protein biofilm interaction network map.

The laboratory was recently awarded a four-year American Cancer Society (ACS) Research Scholar Award in order to ellucidate the role of tyrosine phosphatases in cancers of the immune system. (10/2007)

Graduate student Breann Brown received an NIH F31 NRSA award. (9/2008)

Graduate student David Critton's publication Structural Basis of Substrate Recognition by HePTP was selected as a highlighted manuscript in the journal Biochemistry. (12/2008)

 

 

Wolfgang Peti, Ph.D., Brown University.

Chistopher Seto, Ph.D., Brown University.

Lutz Tautz, Ph.D., The Burnham Institute.

Thomas Woods, Ph.D, Texas A&M University.

Paul Lombroso, M.D., Yale University.

The production of diffraction quality crystals is still one of the time-limiting steps in x-ray crystallography. We use a parallel, rather than serial, approach to clone, express, purify and crystallize multiple constructs of our targets, both alone and as complexes with their binding partners. To do this, we use methods developed by us and others for the high-throughput, parallel cloning, expression, purification, crystallization of multiple protein constructs. This enables us to rapidly identify those constructs and especially complexes most suitable for crystallographic studies.