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Isolation and Crystallization of Heterotrimeric Translation Initiation Factor 2 from Sulfolobus solfataricus


E. A. Stolboushkina*, O. S. Nikonov, and M. B. Garber

Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia; fax: (495) 632-7871; E-mail: esmail@vega.protres.ru

* To whom correspondence should be addressed.

Received March 27, 2008; Revision received April 17, 2008
The structure of the intact heterotrimeric translation initiation factor 2 (e/aIF2) is of great interest due to its key role in the initiator tRNA delivery to the ribosome and in translation initiation regulation in eukaryotes and archaea. We have chosen aIF2 from the hyperthermophilic archaeobacterium Sulfolobus solfataricus (SsoIF2) as an object for crystallization and structural investigations. Genes of the SsoIF2 subunits α, β, and γ were cloned and superexpressed. A method for heterotrimer SsoIF2αβγ purification was elaborated with at least 95% purity. Highly ordered crystals of the full-sized SsoIF2, reflecting X-rays at the resolution up to 2.8 Å, were obtained for the first time.
KEY WORDS: translation initiation, e/aIF2, heterotrimer, Sulfolobus solfataricus, SsoIF2 crystals

DOI: 10.1134/S0006297909010088