title: Bacterial synthesis, purification, and solubilization of membrane protein KCNE3, a regulator of voltage-gated potassium channels.
abstract: An efficient method is described for production of membrane protein KCNE3 and its isotope labeled derivatives ((15)N-, (15)N-/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 alpha-helical conformation. The existence of cross-peaks for all glycines of the (15)N-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.
authors: Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia. goncharuk@nmr.ru
authors: S A
authors: SA
authors: Goncharuk
authors: A A
authors: AA
authors: Shulga
authors: Ya S
authors: YS
authors: Ermolyuk
authors: P K
authors: PK
authors: Kuzmichev
authors: V A
authors: VA
authors: Sobol
authors: E V
authors: EV
authors: Bocharov
authors: V V
authors: VV
authors: Chupin
authors: A S
authors: AS
authors: Arseniev
authors: M P
authors: MP
authors: Kirpichnikov
mesh_terms: D002942:Circular Dichroism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D024642:Potassium Channels, Voltage-Gated / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: C418492:KCNE3 protein, human; D008823:Micelles; D024642:Potassium Channels, Voltage-Gated; D011994:Recombinant Proteins