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A. Kanda, A. Estrada-Cuzcano, D. Hajkova, M. Miyagi, A. Swaroop; Biochemical and Functional Studies of ARMS2, A Mitochondrial Protein Strongly Associated With Susceptibility to Age-Related Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2335.
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© ARVO (1962-2015); The Authors (2016-present)
We and others previously demonstrated that SNP rs10490924 within the ARMS2/LOC387715 gene at 10q26 is strongly associated with susceptibility to age-related macular degeneration and that 12 kDa ARMS2 protein is localized to the mitochondrial outer membrane (Kanda A et al. 2007 PNAS; Kanda A et al. 2008 ARVO). The goal of these studies was to gain additional insights into the function of ARMS2 and its association with AMD.
We performed yeast two-hybrid screening using human retina cDNA library and pulldown assay to identify ARMS2-interacting proteins. The interaction between ARMS2 and candidate proteins was validated by co-immunoprecipitaion followed by immunoblot analysis. We also produced transgenic mice expressing ARMS2 and HTRA1 in photoreceptors.
The yeast two hybrid analysis identified translocase of outer mitochondrial membrane 40 homolog (TOMM40) among others. This interaction was confirmed by co-immunoprecipitation from transfected COS-7 cells. Knockdown of TOMM40 by specific siRNA mislocalized ARMS2. We also identified specific ARMS2-interacting heat shock proteins by mass-spectrometric analysis of proteins obtained by GST-pulldown assay. Moreover, we created Crxp-ARMS2 and Crxp-HTRA1 transgenic mice to examine their role in the retina; their analysis is in progress.
Our data further confirms that ARMS2 is a mitochondrial protein and that TOMM40 is essential for its localization. A better understanding of ARMS2 function will be critical for delineating its involvement in AMD pathogenesis.
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