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This Article Summary Full Text Full Text (PDF) All Versions of this Article: fj.06-6306fjev1 20/14/2636    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Gueorguieva, D. Articles by Pandey, S. Search for Related Content PubMed PubMed Citation Articles by Gueorguieva, D. Articles by Pandey, S.

Identification of single-domain, Bax-specific intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis

Deyzi Gueorguieva^*, Shenghua Li, Nicole Walsh^*, Amit Mukerji^*, Jamshid Tanha and Siyaram Pandey^*,1

^* Department of Chemistry and Biochemistry, University of Windsor, Windsor, Ontario, Canada; and

Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada

¹Correspondence: Department of Chemistry and Biochemistry, University of Windsor, 401 Sunset Ave., Windsor, Ontario, Canada, N9B 3P4. E-mail: spandey{at}uwindsor.ca

ABSTRACT

Bax is a proapoptotic protein implicated in cell death involved in several neurodegenerative diseases. Intracellularly expressed antibody (Ab) fragments (intrabodies) inhibiting Bax function would have potential for developing therapeutics for the aforementioned diseases and can serve as research tools. We report identification, cloning, and functional characterization of several Bax-specific single-domain antibodies (sdAbs). These minimal size Ab fragments, which were isolated from a llama V_HH phage display library by panning, inhibited Bax function in in vitro assays. Importantly, as intrabodies, these sdAbs, which were stably expressed in mammalian cells, were nontoxic to their host cells and rendered them highly resistant to oxidative-stress-induced apoptosis. The intrabodies prevented mitochondrial membrane potential collapse and apoptosis after oxidative stress in the host cells. These anti-Bax V_HHs could be used as tools for studying the role of Bax in oxidative-stress-induced apoptosis and for developing novel therapeutics for the degenerative diseases involving oxidative stress. —Gueorguieva, D., Li, S., Walsh, N., Mukerji, A., Tanha, J., Pandey, S. Identification of single-domain, Bax-specific intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis.