Bacterial synthesis, purification, and solubilization of membrane protein KCNE3, a regulator of voltage-gated potassium channels

S. A. Goncharuk, A. A. Shulga, Ya S. Ermolyuk, P. K. Kuzmichev, V. A. Sobol, E. V. Bocharov, V. V. Chupin, A. S. Arseniev, M. P. Kirpichnikov

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

An efficient method is described for production of membrane protein KCNE3 and its isotope labeled derivatives (15N-, 15N-/ 13C-) in amounts sufficient for structural-functional investigations. The purified protein preparation within different detergent micelles was characterized using dynamic light scattering, CD spectroscopy, and NMR spectroscopy. It is shown that within DPC/LDAO micelles the protein is in monomeric form and acquires mainly α-helical conformation. The existence of cross-peaks for all glycines of the 15N-HSQC NMR spectra as well as relatively small line widths (∼20 Hz) confirm the high quality of the preparation and the possibility of obtaining structural-dynamic information on KCNE3 by high resolution heteronuclear NMR spectroscopy.

Original languageEnglish
Pages (from-to)1344-1349
Number of pages6
JournalBiochemistry (Moscow)
Volume74
Issue number12
DOIs
Publication statusPublished - Dec 2009
Externally publishedYes

Keywords

  • Dynamic light scattering
  • Expression
  • KCNE (MiRP)
  • Membrane protein
  • NMR
  • Purification

ASJC Scopus subject areas

  • Biochemistry

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