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http://dx.doi.org/10.5352/JLS.2007.17.1.018

High-level production and initial crystallization of a Fe65 PTB domain  

Ro, Seung-Hyun (Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota)
Ha, Nam-Chul (College Pharmacy and Research Institute for Drug Development, Pusan National University)
Publication Information
Journal of Life Science / v.17, no.1, 2007 , pp. 18-23 More about this Journal
Abstract
Fe65, a neuron-specific adaptor protein, has two phosphotyrosine binding (PTB) domains. The second PTB (PTB2) domain interacts with intracellular domain fragment (AICD) of amyloid beta precursor protein (APP). Recent studies suggested that tile complex is composed of AICD and Fe65 transactivates genes that are responsible for neuronal cell death in Alzheimer's disease (AD). Therefore, a compound inhibiting the interaction between Fe65 and AICD can be a drug candidate to treat AD. However, it remains unclear how Fe65 recognizes AICD at a molecular level. Here, we report high-level production of the PTB2 domain of Fe65 in the baculovirus system. We found that the baculovirus system is an efficient method to obtain the Fe65 PTB2 domain, compared with the bacterial and mammalian expression systems. The purified recombinant protein was used for crystallization to determine its crystal structure helping to understand the molecular mechanism of Fe65-dependent signaling and to design its inhibitors.
Keywords
Alzheimer's disease; crystallization; overexpression; Fe65;
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