In vitro synthesis, tetramerization and single channel characterization of virus-encoded potassium channel Kcv

Jiwook Shim, Mingming Yang, Li Qun Gu

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Chlorella virus-encoded membrane protein Kcv represents a new class of potassium channel. This 94-amino acids miniature K+ channel consists of two trans-membrane α-helix domains intermediated by a pore domain that contains a highly conserved K+ selectivity filter. Therefore, as an archetypal K+ channel, the study of Kcv may yield valuable insights into the structure-function relationships underlying this important class of ion channel. Here, we report a series of new properties of Kcv. We first verified Kcv can be synthesized in vitro. By co-synthesis and assembly of wild-type and the tagged version of Kcv, we were able to demonstrate a tetrameric stoichiometry, a molecular structure adopted by all known K+ channels. Most notably, the tetrameric Kcv complex retains its functional integrity in SDS (strong detergent)-containing solutions, a useful feature that allows for direct purification of protein from polyacrylamide gel. Once purified, the tetramer can form single potassium-selective ion channels in a lipid bilayer with functions consistent to the heterologously expressed Kcv. These finding suggest that the synthetic Kcv can serve as a model of virus-encoded K+ channels; and its newly identified properties can be applied to the future study on structure-determined mechanisms such as K+ channel functional stoichiometry.

Original languageEnglish (US)
Pages (from-to)1027-1034
Number of pages8
JournalFEBS Letters
Volume581
Issue number5
DOIs
StatePublished - Mar 6 2007

Fingerprint

Potassium Channels
Ion Channels
Viruses
Stoichiometry
Chlorella
Lipid bilayers
Lipid Bilayers
Molecular Structure
Detergents
Molecular structure
Purification
Potassium
Membrane Proteins
Membranes
Amino Acids
Proteins
In Vitro Techniques
polyacrylamide gels

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

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abstract = "Chlorella virus-encoded membrane protein Kcv represents a new class of potassium channel. This 94-amino acids miniature K+ channel consists of two trans-membrane α-helix domains intermediated by a pore domain that contains a highly conserved K+ selectivity filter. Therefore, as an archetypal K+ channel, the study of Kcv may yield valuable insights into the structure-function relationships underlying this important class of ion channel. Here, we report a series of new properties of Kcv. We first verified Kcv can be synthesized in vitro. By co-synthesis and assembly of wild-type and the tagged version of Kcv, we were able to demonstrate a tetrameric stoichiometry, a molecular structure adopted by all known K+ channels. Most notably, the tetrameric Kcv complex retains its functional integrity in SDS (strong detergent)-containing solutions, a useful feature that allows for direct purification of protein from polyacrylamide gel. Once purified, the tetramer can form single potassium-selective ion channels in a lipid bilayer with functions consistent to the heterologously expressed Kcv. These finding suggest that the synthetic Kcv can serve as a model of virus-encoded K+ channels; and its newly identified properties can be applied to the future study on structure-determined mechanisms such as K+ channel functional stoichiometry.",
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In vitro synthesis, tetramerization and single channel characterization of virus-encoded potassium channel Kcv. / Shim, Jiwook; Yang, Mingming; Gu, Li Qun.

In: FEBS Letters, Vol. 581, No. 5, 06.03.2007, p. 1027-1034.

Research output: Contribution to journalArticle

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