Aggregatibacter actinomycetemcomitans is a gram negative pathogen that is the etiologic agent of localized aggressive periodontitis (LAP) and other systemic infections, including infective endocarditis. LAP, a disease which affects medically-underserved children in both the United States and throughout the world, is characterized by a breakdown of the periodontal ligament and alveolar bone structure that holds the teeth in place. Without intervention, loss of teeth occurs, causing both a cosmetic deformity and a functional defect. The manner by which A. actinomycetemcomitans causes disease has not been determined;however, it is known that the bacterium produces several putative virulence factors, including a leukotoxin (LtxA) that is a member of the repeats-in-toxin (RTX) family. This toxin kills human white blood cells and likely plays a key role in the ability of the organism to evade the immune response during its establishment of an ecological niche in the host. Thus, preventing or interfering with LtxA activity may be one option for successful treatment of the disease. The mechanism by which LtxA exerts its lethal effect on the host cell involves the formation of a physical disruption, such as a pore. Specifically, the interaction of the toxin and membrane results in a bending of the bilayer that leads to the formation of a toroidal, or lipid-lined, pore. A logical extension of this argument would suggest that LtxA will therefore interact most strongly with lipids that are able to bend or curve more easily. The proposed study is designed to explore the ability of LtxA to damage cells by altering the structure of their membranes. In the first aim, the effect of lipid curvature on LtxA-induced membrane changes, such as phase behavior and pore formation, will be characterized. In the second aim, the effect of lipid curvature on LtxA structure will be determined and the relation between those LtxA structural changes and the membrane changes observed in Aim 1 will be examined.
The final aim will elucidate the biophysical properties of LtxA- induced pores. This study will allow determination of the role of the physical properties of the membrane, specifically curvature, on LtxA-induced membrane disruption, and will establish the mechanism of this disruption.

Public Health Relevance

Periodontal diseases are infections caused by bacteria that reside in dental plaque under the gums. To colonize this area, bacteria must produce factors (specific proteins) that enable them to survive in this harsh environment and destroy the white blood cells that protect the host from this bacterial onslaught. The question asked in this study is how one of these proteins, a toxin, helps the bacteria to survive by killing the host white blood cells.

National Institute of Health (NIH)
National Institute of Dental & Craniofacial Research (NIDCR)
Postdoctoral Individual National Research Service Award (F32)
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NIDCR Special Grants Review Committee (DSR)
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Frieden, Leslie A
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University of Pennsylvania
Schools of Dentistry
United States
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Koufos, Evan; Chang, En Hyung; Rasti, Elnaz S et al. (2016) Use of a Cholesterol Recognition Amino Acid Consensus Peptide To Inhibit Binding of a Bacterial Toxin to Cholesterol. Biochemistry 55:4787-97
Brown, Angela C; Balashova, Nataliya V; Epand, Richard M et al. (2013) Aggregatibacter actinomycetemcomitans leukotoxin utilizes a cholesterol recognition/amino acid consensus site for membrane association. J Biol Chem 288:23607-21
Walters, M J; Brown, A C; Edrington, T C et al. (2013) Membrane association and destabilization by Aggregatibacter actinomycetemcomitans leukotoxin requires changes in secondary structures. Mol Oral Microbiol 28:342-53
Brown, Angela C; Boesze-Battaglia, Kathleen; Du, Yurong et al. (2012) Aggregatibacter actinomycetemcomitans leukotoxin cytotoxicity occurs through bilayer destabilization. Cell Microbiol 14:869-81
Fong, K P; Tang, H-Y; Brown, A C et al. (2011) Aggregatibacter actinomycetemcomitans leukotoxin is post-translationally modified by addition of either saturated or hydroxylated fatty acyl chains. Mol Oral Microbiol 26:262-76