The nature of protein-protein interactions of crystallins forms the basis for understanding the transparent and opaque properties of the normal and cataractous lens. Previous physical studies have suggested that weak, noncovalent interactions of these lens crystallins (i.e. alpha-gamma, alpha-beta, gamma-beta) may exist in the lens, where they are necessary for lens transparency. These interactions have hitherto not been well characterized, since they involve interactions that dissociate under normal analytical conditions. The central hypothesis of this proposal is that under true equilibrium conditions, noncovalent interactions of alpha-gamma, alpha-beta, and beta-gamma crystallins exist, and because of posttranslational modifications or genetic mutations, these interactions are perturbed during cataractogenesis of the lens, resulting in loss of lens transparency. We have developed the technique of microequilibrium dialysis to both detect and quantitate possible interactions.
The specific aim of this proposal will be to use this methodology to quantitate noncovalent interactions between different crystallins during cataractogenesis of the human lens. Using these novel methods, we will be able to ascertain the existence of these interactions and their possible alterations during lens opacification.

Public Health Relevance

of this research to public health: Opacification of the lens, known as cataract, is the leading cause of blindness in the world. Opacification is thought to be caused by abnormal interactions of proteins in the lens. The proposed research will characterize these interactions, and determine how they are altered during formation of cataracts.

National Institute of Health (NIH)
National Eye Institute (NEI)
Research Project (R01)
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Anterior Eye Disease Study Section (AED)
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Araj, Houmam H
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Kansas State University
Schools of Arts and Sciences
United States
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Takemoto, Larry; Sorensen, Christopher M (2008) Protein-protein interactions and lens transparency. Exp Eye Res 87:496-501
Sabah, Judith R; Schultz, Bruce D; Brown, Zach W et al. (2007) Transcytotic passage of albumin through lens epithelial cells. Invest Ophthalmol Vis Sci 48:1237-44
Brown, Zachery; Ponce, Aldo; Lampi, Kirsten et al. (2007) Differential binding of mutant (R116C) and wildtype alphaA crystallin to actin. Curr Eye Res 32:1051-4
Takemoto, Larry J; Ponce, Aldo A (2006) Decreased association of aged alpha crystallins with gamma crystallins. Exp Eye Res 83:793-7
Lin, Dingbo; Barnett, Micheal; Grauer, Laura et al. (2005) Expression of superoxide dismutase in whole lens prevents cataract formation. Mol Vis 11:853-8
Ponce, Aldo; Takemoto, Larry (2005) Screening of crystallin-crystallin interactions using microequilibrium dialysis. Mol Vis 11:752-7
Peterson, James J; Young, Malin M; Takemoto, Larry J (2004) Probing alpha-crystallin structure using chemical cross-linkers and mass spectrometry. Mol Vis 10:857-66
Nguyen, Thu Annelise; Takemoto, Larry J; Takemoto, Dolores J (2004) Inhibition of gap junction activity through the release of the C1B domain of protein kinase Cgamma (PKCgamma) from 14-3-3: identification of PKCgamma-binding sites. J Biol Chem 279:52714-25
Boyle, Daniel L; Takemoto, Larry; Brady, James P et al. (2003) Morphological characterization of the Alpha A- and Alpha B-crystallin double knockout mouse lens. BMC Ophthalmol 3:3
Reddy, V N; Giblin, F J; Lin, L R et al. (2001) Glutathione peroxidase-1 deficiency leads to increased nuclear light scattering, membrane damage, and cataract formation in gene-knockout mice. Invest Ophthalmol Vis Sci 42:3247-55

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