The role of extramembranous cytoplasmic termini in assembly and stability of the tetrameric K(+)-channel KcsA.
Membrane-active alcohol 2,2,2-trifluoroethanol has been proven to be an attractive tool in the investigation of the intrinsic stability of integral membrane protein complexes by taking K(+)-channel KcsA as a suitable and representative ion channel. In the present study, the roles of both cytoplasmic N and C termini in channel assembly and stability of KcsA were determined. The N terminus (1-18 residues) slightly increased tetramer stability via electrostatic interactions in the presence of 30 mol.% acidic phosphatidylglycerol (PG) in phosphatidylcholine lipid bilayer. Furthermore, the N terminus was found to be potentially required for efficient channel (re)assembly. In contrast, truncation of the C terminus (125-160 residues) greatly facilitated channel reversibility from either a partially or a completely unfolded state, and this domain was substantially involved in stabilizing the tetramer in either the presence or absence of PG in lipid bilayer. These studies provide new insights into how extramembranous parts play their crucial roles in the assembly and stability of integral membrane protein complexes.
Raja
Mobeen
M
Department of Biochemistry of Membranes, Centre for Biomembranes and Lipid Enzymology, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands. mraja@pmcol.ualberta.ca
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D008051:Lipid Bilayers / Q000737:chemistry; D010715:Phosphatidylglycerols; D015221:Potassium Channels / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D017510:Protein Folding; D055503:Protein Multimerization; D055550:Protein Stability; D013050:Spectrometry, Fluorescence; D014270:Trifluoroethanol / Q000494:pharmacology; D014364:Tryptophan / Q000737:chemistry
D001426:Bacterial Proteins; D008051:Lipid Bilayers; D010715:Phosphatidylglycerols; D015221:Potassium Channels; C107790:prokaryotic potassium channel; D014270:Trifluoroethanol; D014364:Tryptophan