Chlorella virus ATCV-1 encodes a functional potassium channel of 82 amino acids

Biochem J. 2009 May 13;420(2):295-303. doi: 10.1042/BJ20090095.

Abstract

Chlorella virus PBCV-1 (Paramecium bursaria chlorella virus-1) encodes the smallest protein (94 amino acids, named Kcv) previously known to form a functional K+ channel in heterologous systems. In this paper, we characterize another chlorella virus encoded K+ channel protein (82 amino acids, named ATCV-1 Kcv) that forms a functional channel in Xenopus oocytes and rescues Saccharomyces cerevisiae mutants that lack endogenous K+ uptake systems. Compared with the larger PBCV-1 Kcv, ATCV-1 Kcv lacks a cytoplasmic N-terminus and has a reduced number of charged amino acids in its turret domain. Despite these deficiencies, ATCV-1 Kcv accomplishes all the major features of K+ channels: it assembles into a tetramer, is K+ selective and is inhibited by the canonical K+ channel blockers, barium and caesium. Single channel analyses reveal a stochastic gating behaviour and a voltage-dependent conductance that resembles the macroscopic I/V relationship. One difference between PBCV-1 and ATCV-1 Kcv is that the latter is more permeable to K+ than Rb+. This difference is partially explained by the presence of a tyrosine residue in the selective filter of ATCV-1 Kcv, whereas PBCV-1 Kcv has a phenylalanine. Hence, ATCV-1 Kcv is the smallest protein to form a K+ channel and it will serve as a model for studying structure-function correlations inside the potassium channel pore.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Barium / pharmacology
  • Cation Transport Proteins / genetics
  • Cation Transport Proteins / physiology
  • Chlorella / virology*
  • Female
  • Genetic Complementation Test
  • Ion Transport
  • Membrane Potentials / drug effects
  • Molecular Sequence Data
  • Mutation
  • Oocytes / metabolism
  • Oocytes / physiology
  • Patch-Clamp Techniques
  • Potassium / metabolism
  • Potassium Channel Blockers / pharmacology
  • Potassium Channels / chemistry
  • Potassium Channels / genetics
  • Potassium Channels / physiology*
  • Protein Structure, Secondary
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / physiology
  • Sequence Homology, Amino Acid
  • Viral Proteins / chemistry
  • Viral Proteins / genetics
  • Viral Proteins / physiology*
  • Xenopus laevis

Substances

  • Cation Transport Proteins
  • Kcv potassium channel, Chlorella virus
  • Potassium Channel Blockers
  • Potassium Channels
  • Saccharomyces cerevisiae Proteins
  • TRK2 protein, S cerevisiae
  • Viral Proteins
  • TRK1 protein, S cerevisiae
  • Barium
  • Potassium