Abstract

Expression of G-Protein Coupled Receptors in Escherichia coli for Structural Studies

L. E. Petrovskaya^1*, A. A. Shulga¹, O. V. Bocharova¹, Ya. S. Ermolyuk¹, E. A. Kryukova¹, V. V. Chupin¹, M. J. J. Blommers², A. S. Arseniev¹, and M. P. Kirpichnikov^1,3

¹Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997 Moscow, Russia; fax: (495) 330-6983; E-mail: lpetr65@yahoo.com

²Novartis Institutes for BioMedical Research, Basel, Switzerland

³Faculty of Biology, Lomonosov Moscow State University, 119991 Moscow, Russia

^* To whom correspondence should be addressed.

Received December 12, 2009; Revision received February 22, 2010
To elaborate a high-performance system for expression of genes of G-protein coupled receptors (GPCR), methods of direct and hybrid expression of 17 GPCR genes in Escherichia coli and selection of strains and bacteria cultivation conditions were investigated. It was established that expression of most of the target GPCR fused with the N-terminal fragment of OmpF or Mistic using media for autoinduction provides high output (up to 50 mg/liter).
KEY WORDS: autoinduction, expression in E. coli, GPCR, OmpF, Mistic
DOI: 10.1134/S0006297910070102

Expression of G-Protein Coupled Receptors in Escherichia coli for Structural Studies

L. E. Petrovskaya1*, A. A. Shulga1, O. V. Bocharova1, Ya. S. Ermolyuk1, E. A. Kryukova1, V. V. Chupin1, M. J. J. Blommers2, A. S. Arseniev1, and M. P. Kirpichnikov1,3

L. E. Petrovskaya^1*, A. A. Shulga¹, O. V. Bocharova¹, Ya. S. Ermolyuk¹, E. A. Kryukova¹, V. V. Chupin¹, M. J. J. Blommers², A. S. Arseniev¹, and M. P. Kirpichnikov^1,3