Transparent cellular structure is the fundamental property of the biological lens. The next five year application will investigate the molecular basis for the transparent structure of lens cells. The PI's past studies of the collective interactions measured by Tc led to a clinical trial of pantethine as an anticataract agent in humans. Pantethine was used because it was effective against protein aggregation and lens opacification in animal models. During the next five years, the functional motifs of selected human lens proteins that appear to be involved with the interactions important for development and maintenance of lens cell transparency will be investigated. Recombinant human lens crystallins (wild-type and mutant) will be expressed, purified and characterized using biochemical assays for interactions involving human crystallins (AIM 1). Novel quantitative methods for Fourier, power law and fractal analysis will be used to characterize the microscopic structure of lens cells and of model solutions of human lens cytoplasmic proteins (AIM 2). The methods will be used to investigate and quantify the size and order/disorder of major structural components found in lens cytoplasm and will evaluate the fractal dimension of lens cell structure. Preliminary results suggest that alpha B crystallin and cytoskeletal proteins have important functions in the development and maintenance of lens cell transparency. Transgenic and knockout models for cataract will be used for in vivo studies of the relationships between lens crystallins, cytoskeletal proteins and lens opacification (AIM 3). The results of the proposed studies will have direct application in the design and testing of new therapeutic approaches to inhibit protein aggregation and lens opacification in humans.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY004542-19
Application #
6178412
Study Section
Visual Sciences A Study Section (VISA)
Program Officer
Liberman, Ellen S
Project Start
1982-06-01
Project End
2003-05-31
Budget Start
2000-06-01
Budget End
2001-05-31
Support Year
19
Fiscal Year
2000
Total Cost
$340,721
Indirect Cost
Name
University of Washington
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Clark, John I (2016) Functional sequences in human alphaB crystallin. Biochim Biophys Acta 1860:240-5
Anderson, David M G; Floyd, Kyle A; Barnes, Stephen et al. (2015) A method to prevent protein delocalization in imaging mass spectrometry of non-adherent tissues: application to small vertebrate lens imaging. Anal Bioanal Chem 407:2311-20
Clark, John I (2013) Self-assembly of protein aggregates in ageing disorders: the lens and cataract model. Philos Trans R Soc Lond B Biol Sci 368:20120104
Gokhin, David S; Nowak, Roberta B; Kim, Nancy E et al. (2012) Tmod1 and CP49 synergize to control the fiber cell geometry, transparency, and mechanical stiffness of the mouse lens. PLoS One 7:e48734
Clark, Tyler J W; Houck, Scott A; Clark, John I (2012) Hemoglobin interactions with ?B crystallin: a direct test of sensitivity to protein instability. PLoS One 7:e40486
Greiling, Teri M S; Clark, John I (2012) New insights into the mechanism of lens development using zebra fish. Int Rev Cell Mol Biol 296:1-61
Qu, Bo; Landsbury, Andrew; Schönthaler, Helia Berrit et al. (2012) Evolution of the vertebrate beaded filament protein, Bfsp2; comparing the in vitro assembly properties of a ""tailed"" zebrafish Bfsp2 to its ""tailless"" human orthologue. Exp Eye Res 94:192-202
Houck, Scott A; Landsbury, Andrew; Clark, John I et al. (2011) Multiple sites in ?B-crystallin modulate its interactions with desmin filaments assembled in vitro. PLoS One 6:e25859
Houck, Scott A; Clark, John I (2010) Dynamic subunit exchange and the regulation of microtubule assembly by the stress response protein human alphaB crystallin. PLoS One 5:e11795
Greiling, Teri M S; Aose, Masamoto; Clark, John I (2010) Cell fate and differentiation of the developing ocular lens. Invest Ophthalmol Vis Sci 51:1540-6

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