Jed N Lampe, PhD

Assistant Professor
Ph.D., University of Washington, 2007


Research Interests:

In the Lampe laboratory, we combine powerful tools from both bioanalytical and biophysical chemistry to understand structure-function relationships in protein drug targets and to design new chemical entities to modulate these targets.  Currently, there are two major research initiatives in our lab:


Understanding the role of cytochrome P450 CYP46A1 in traumatic brain injury

Traumatic brain injury (TBI) is a major cause of long-term disability in the United States, affecting at least 1.7 million Americans each year and is a contributing factor in 30% of all injury-related deaths (U.S. Department of Health and Human Services, 2010). Failure to diagnose TBI correctly after injury can lead to ineffective treatment and negative long-term outcomes. Accordingly, there is a critical need for both improved diagnostic tests that can determine the presence and severity of TBI, and for a better understanding of the underlying pathophysiology of TBI. The human brain is composed of ~ 20% cholesterol, the synthesis and degradation of which has been demonstrated to be important for long-term memory and learning, and may also be important in recovery from TBI. The cytochrome P450 CYP46A1 is a neural specific monoxygenase enzyme that is responsible for cholesterol turnover in the brain, producing the oxidized metabolite 24(S)-hydroxycholesterol from cholesterol. Preliminary studies from our lab and others suggest that circulating plasma levels of 24(S)-hydroxycholesterol are elevated post-TBI in rat models. This metabolite has been demonstrated to effect expression of genes involved in cholesterol synthesis and degradation, which may be important in the brain's response to TBI. Additionally, elevated plasma levels of 24(S)-hydroxycholesterol may serve as a peripheral biomarker for the diagnosis of TBI. Our long-term goal for this project is to develop novel tools for the diagnosis and treatment of TBI through investigations that will clarify the role of CYP46A1 and the cholesterol metabolites it produces in the underlying biochemical pathophysiology of TBI.


Designing novel inhibitors to target the anti-apoptotic protein Survivin

Survivin is an essential protein for both cell division and the inhibition of apoptosis. Moreover, its expression is upregulated in nearly all cancers. Inhibition of survivin expression has been shown to inhibit tumor growth and increase survival rates in several neoplastic animal models. Survivin carries out its important cellular functions through interactions with numerous protein partners in the cell. During mitosis, the survivin protein associates with Borealin and INCENP to form the chromosomal passenger complex, which is essential to chromosome segregation and mitotic spindle formation. Similarly, survivin prevents apoptosis in cancer cells by inhibiting cellular caspase activity, which can be blocked when survivin associates with SMAC/Diablo. The current focus of this project is to develop high-throughput fluorescence assays for screening small molecule libraries in order to identify lead compounds that may disrupt survivin's interactions with its protein partners. Using both a unique "click chemistry" mediated approach and more traditional fluorescence polarization technology, we intend to screen focused small molecule libraries for inhibition of both the survivin/SMAC and the survivin/Borealin interaction. Proper lead identification will then allow us optimize candidates further to improve affinity and specificity using standard medicinal chemistry techniques. Prospective drug candidates will be tested with in vitro cell culture and in vivo mouse model systems. Targeted disruption of survivin protein-protein interactions is likely to be a fruitful approach to treat currently intractable cancers.



Selected Publications:

Varfaj, F., Lampe, J.N., Ortiz de Montellano, P.R. (2012). Role of Cysteine Residues in Heme Binding to Human Heme Oxygenase-2 Elucidated by 2D NMR Spectroscopy. J. Biol. Chem. 287, 35181-35191.

Williams, C.D., Koerner, M.R., Lampe, J.N., Farhood, A., Jaeschke, H. (2011). Mouse strain-dependent caspase activation during acetaminophen hepatotoxicity does not result in apoptosis or modulation of inflammation. Toxicol. Appl. Pharmacol. 257, 449-58.

Brandman, R., Lampe, J.N., Brandman, Y., Ortiz de Montellano, P.R. (2011). Active-site residues move independently from the rest of the protein in a 200ns molecular dynamics simulation of cytochrome P450 CYP119. Arch. Biochem. Biophys. 506, 9594-9603.

Lampe, J.N., Brandman, R., Sivaramakrishnan, S., and Ortiz de Montellano, P. R. (2010). Two-dimensional NMR and all-atom molecular dynamics of cytochrome P450 CYP119 reveal hidden conformational substates. J. Biol. Chem. 285, 9594-9603.

Lampe, J.N., Floor, S. N., Gross, J. D., Nishida, C. R., Jiang, Y., Trnka, M. J., and Ortiz de Montellano, P. R. (2008). Ligand-induced conformational heterogeneity of cytochrome P450 CYP119 identified by 2D NMR spectroscopy with the unnatural amino acid 13C-p -methoxyphenylalanine. J. Am. Chem. Soc., 130, 16168-16169.

Nath, A., Fernandez, C., Lampe, J.N., and Atkins, W. M. (2008). Spectral resolution of a second binding site for Nile Red on cytochrome P4503A4. Arch. Biochem. Biophys. 474, 198-204.

Lampe, J.N., Fernandez, C., Nath, A., and Atkins, W.M. (2007). Nile Red is a fluorescent allosteric substrate of cytochrome P450 3A4. Biochemistry 47, 509-516.

Lampe, J.N., and Atkins, W.M. (2006). Time-resolved fluorescence studies of heterotropic ligand binding to cytochrome P450 3A4. Biochemistry 45, 12204-12215.

Wen, B., Lampe, J.N., Roberts, A.G., Atkins, W.M., David Rodrigues, A., and Nelson, S.D. (2006). Cysteine 98 in CYP3A4 contributes to conformational integrity required for P450 interaction with CYP reductase. Arch. Biochem. Biophys. 454, 42-54.

Roberts, A.G., Diaz, M.D., Lampe, J.N., Shireman, L.M., Grinstead, J.S., Dabrowski, M.J., Pearson, J.T., Bowman, M.K., Atkins, W.M., and Campbell, A.P. (2006). NMR studies of ligand binding to P450(eryF) provides insight into the mechanism of cooperativity. Biochemistry 45, 1673-1684.

Lampe, J.N., and Atkins, W.M. (2006). Time-resolved fluorescence studies of heterotropic ligand binding to cytochrome P450 3A4. Biochemistry 45, 12204-12215.

Wen, B., Doneanu, C.E., Lampe, J.N., Roberts, A.G., Atkins, W.M., and Nelson, S.D. (2005). Probing the CYP3A4 active site by cysteine scanning mutagenesis and photoaffinity labeling. Arch. Biochem. Biophys. 444, 100-111.

Compagno, D., Lampe, J.N., Bourget, C., Kutyavin, I.V., Yurchenko, L., Lukhtanov, E.A., Gorn, V.V., Gamper, H.B., Jr., and Toulme, J.J. (1999). Antisense oligonucleotides containing modified bases inhibit in vitro translation of Leishmania amazonensis mRNAs by invading the mini-exon hairpin. J. Biol. Chem. 274, 8191-8198.

Lampe, J.N., Kutyavin, I.V., Rhinehart, R., Reed, M.W., Meyer, R.B., and Gamper, H.B., Jr. (1997). Factors influencing the extent and selectivity of alkylation within triplexes by reactive G/A motif oligonucleotides. Nucleic Acids Res. 25, 4123-4131.

Last modified: Sep 17, 2013

jlampe

Contact

Jed N Lampe, PhD
Assistant Professor

4069 HLSIC; MS-1018
3901 Rainbow Blvd.
Kansas City, Kansas 66160

P: (913) 588-4760
F: (913) 588-7501
jlampe@kumc.edu

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