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Harry LeVine, III, Ph.D. Research Publications | Publications | Lab | PubMed Misfolded, aberrantly polymerized proteins are a hallmark of a number of neurodegenerative and other diseases. Those impacting the nervous system include Alzheimer's disease, the tauopathies (e.g. Pick;’s disease), polyglutamine diseases (e.g. Huntington's disease), amyotrophic lateral sclerosis, the prion diseases (i.e.. variant CJD ; “mad cow“ disease), and the cerebral amyloid angiopathies. Eighteen additional different disease-associated proteins and peptides are officially recognized as amyloids - insoluble fibrillar forms of normally soluble proteins that have adopted cross-β-sheet secondary structures. In all of these cases, a specific, conformationally unstable protein with a strong tendency to self-associate assembles into characteristic insoluble, protease-resistant structures. In the familial diseases, point-mutations in the encoding genes destabilize the normal folded proteins, increasing their tendency to polymerize and raising the odds that disease will appear. Most of these diseases occur in mid- to late life, although certain mutations can produce early onset. Alzheimer’s disease is characterized histologically by the accumulation of two insoluble proteins, the β-peptide and the microtubule protein tau, in plaque and tangle lesions, respectively. Beginning around age fifty, the β-peptide accumulates in the brain parenchyma, first as histologically invisible soluble forms, and then depositing in visualizable amorphous plaques, some of which are converted to the classic fibrillar senile plaques, possibly by the action of microglia. Genetic evidence from early onset familial forms of the disease implicates the β-peptide as a primary etiologic agent of Alzheimer’s disease. I joined the Biochemistry Department and the Center on Aging at UK after 28 years of research and drug discovery in the pharmaceutical industry at Burroughs-Wellcome, Glaxo, Parke-Davis, and Pfizer. During the early 1990’s I was focused on mechanisms of Alzheimer’s β-peptide production and fibril formation. Alongside this basic research, I was involved in drug discovery efforts, most recently in the search for inhibitors of polymerization of the β-peptide with viable drug properties from in vitro screening through animal models up to the pre-Phase I clinical trial stage. At the University of Kentucky, my laboratory applies biochemical and biophysical approaches to learn how toxic oligomeric forms of the Alzheimer’s β-peptide are generated, and why they persist in the test tube, in cells, in animal models, and in humans. Our goals include a detailed mechanistic understanding of oligomer formation, cellular mechanisms for dealing with intracellular misfolded proteins, and the pharmacological manipulation of these toxic oligomers for therapy. Mechanistic similarities among the protein misfolding diseases suggest that many of the lessons learned with the Alzheimer’s β-peptide will be applicable to the other diseases, and will facilitate the application of pharmaceuticals targeting protein polymerization. Laboratory Projects Chaperone network interactions with oligomeric forms of the
Alzheimer’s b-peptide. _______________________________________________________________________________________________ LeVine, III, H. "Thioflavine T Interaction with Synthetic Alzheimer's Disease β - Amyloid Peptides: Detection of Amyloid Aggregation in Solution", Protein Science 2: 404-410 (1993). LeVine, III, H. "Soluble Multimeric Alzheimer β(1‑40) Pre - Amyloid Complexes in Dilute Solution.", Neurobiology of Aging 16: 755-764 (1995). LeVine, III, H. "Thioflavine T Interaction with Amyloid β-Sheet Structures." Amyloid: The International Journal of Experimental and Clinical Investigation 2: 1-6 (1995). LeVine, III, H. "Stopped-flow Kinetics Reveal Multiple Phases of Thioflavine T Binding to Alzheimer β(1-40)", Arch. Biochem. Biophys. 342: 306-316 (1997). LeVine, III, H. "125I-Labeled ApoE Binds Competitively to b(1-40) Fibrils with Pathological Chaperone Proteins", Amyloid: The International Journal of Experimental and Clinical Investigation, 7: 83-89 (2000). Walker, L. C. and LeVine, III, H. "The cerebral proteopathies" Neurobiology of Aging 21: 559-561 (2000). Wegiel, J., Wang, K.-C., Imaki, H., Rubenstein, R., Wronska, A., Osuchowski, M., Lipinski, WJ., Walker, LC., LeVine, III, H. “The role of microglial cells and astrocytes in fibrillar plaque evolution in transgenic APPsw mice”, Neurobiology of Aging 22: 49-61 (2001). LeVine, III, H. “4, 4’-dianilino-1, 1’-binaphthyl-5, 5’-disulfonate (bis-ANS) Reports on Non-b-Sheet Conformers of Alzheimer’s Peptide b(1-40)”, Arch. Biochem. Biophys. 404: 106-115 (2002). Reviews: LeVine, III, H. "Quantification of b-Sheet Amyloid Fibril Structures with Thioflavin T." Meth. Enzymol. 309: 274-284 (1999). LeVine, III, H. and Scholten, J. D. "Screening for Pharmacologic Inhibitors of Amyloid Fibril Formation." Meth. Enzymol. 309: 467-476 (1999). LeVine, III, H. "The Challenge of Inhibiting Ab Fibrillogenesis (Review) Current Medicinal Chemistry 9(11): 1121-1133 (2002). Walker, L. C. and LeVine, III, H. “The Cerebral Proteopathies: Neurodegenerative Disorders of Protein Conformation and Assembly”, Molecular Neurobiology 21(1/2) 83-95 (2001). Walker, L.C., Bian, F., Callahan, M. J., Lipinski, W. J., Durham, R. A., and LeVine, III, H.. “Modeling Alzheimer’s Disease and Other Proteopathies In Vivo: Is Seeding the Key?, Amino Acids 23: 83-88 (2002).Walker, L. C. and LeVine, III, H. “Proteopathy: The Next Therapeutic Frontier?”, Current Opinion in Investigational Drugs 3(5): 782-787 (2002). Mazur-Kolecka, B., Frackowiak, J., LeVine, III, H., Haske, T., Evans, L., Sukontasup, T., and Golobek, A. “TGFb1 Enhances formation of cellular Ab /ApoE deposits in vascular myocytes”, Neurobiology of Aging, 24: 355-364 (2003). |
