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 language | English (US) |
|---|---|
| Pages (from-to) | 1027-1034 |
| Number of pages | 8 |
| Journal | FEBS Letters |
| Volume | 581 |
| Issue number | 5 |
| DOIs | |
| State | Published - Mar 6 2007 |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology
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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 journal › Article
TY - JOUR
T1 - In vitro synthesis, tetramerization and single channel characterization of virus-encoded potassium channel Kcv
AU - Shim, Jiwook
AU - Yang, Mingming
AU - Gu, Li Qun
PY - 2007/3/6
Y1 - 2007/3/6
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=33847111613&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33847111613&partnerID=8YFLogxK
U2 - 10.1016/j.febslet.2007.02.005
DO - 10.1016/j.febslet.2007.02.005
M3 - Article
C2 - 17316630
AN - SCOPUS:33847111613
VL - 581
SP - 1027
EP - 1034
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 5
ER -