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Recombinant expression, purification and characterization of Kch, a putative Escherichia coli potassium channel protein
Author(s) -
Voges Dieter,
Jap Bing K
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00509-2
Subject(s) - escherichia coli , chemistry , recombinant dna , biochemistry , molecular mass , potassium , microbiology and biotechnology , chromatography , biology , gene , enzyme , organic chemistry
The Escherichia coli gene kch , similar in primary structure to eukaryotic voltage‐gated potassium channels, was cloned and overexpressed in E. coli . The protein was solubilized from the plasma membrane with dodecylmaltopyranoside, lauryldimethylamine oxide or N ‐laurylsarcosine and was purified in milligram amounts by imidazole elution from a nickel‐chelate column. The molecular mass of the purified protein in a number of detergents with 12 carbon atom chains suggests that rKch forms primarily tetramers of the 50 kDa monomers. CD spectroscopy of the purified protein indicates a significant α‐helical content that is preserved upon addition of SDS.