Home LiteratureArticle Details
PMID: 9230130 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Synthetic putative transmembrane region of minimal potassium channel protein (minK) adopts an alpha-helical conformation in phospholipid membranes.

The Biochemical journal ·Vol. 325 ( Pt 2) ·1997-07-15 ·Pages 475-9

Mercer EA, Abbott GW, Brazier SP, Ramesh B, Haris PI, Srai SK

Abstract

Minimal potassium channel protein (minK) is a potassium channel protein consisting of 130 amino acids, possessing just one putative transmembrane domain. In this study we have synthesized a peptide with the amino acid sequence RDDSKLEALYILMVLGFFGFFTLGIMLSYI, containing the putative transmembrane region of minK, and analysed its secondary structure by using Fourier-transform IR and CD spectroscopy. The peptide was virtually insoluble in aqueous buffer, forming intermolecular beta-sheet aggregates. On attempted incorporation of the peptide into phospholipid membranes with a method involving dialysis, the peptide adopted a predominantly intermolecular beta-sheet conformation identical with that of the peptide in aqueous buffer, in agreement with a previous report [Horvàth, Heimburg, Kovachev, Findlay, Hideg and Marsh, (1995) Biochemistry 34, 3893-3898]. However, by using an alternative method of incorporating the peptide into phospholipid membranes we found that the peptide adopted a predominantly alpha-helical conformation, a finding consistent with various proposed structural models. These observed differences in secondary structure are due to artifacts of aggregation of the peptide before incorporation into lipid.

MeSH Terms
Amino Acid Sequence Circular Dichroism Lysophosphatidylcholines/chemistry Membrane Lipids/chemistry Membranes, Artificial Molecular Sequence Data Peptide Fragments/chemical synthesis,chemistry Phospholipids/chemistry Potassium Channels/chemistry Protein Structure, Secondary Spectroscopy, Fourier Transform Infrared
Chemicals
Lysophosphatidylcholines Membrane Lipids Membranes, Artificial Peptide Fragments Phospholipids Potassium Channels
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Mercer E A
Department of Biochemistry and Molecular Biology, Royal Free Hospital School of Medicine, Rowland Hill Street, London NW3 2PF, U.K.
Abbott G W
Brazier S P
Ramesh B
Haris P I
Srai S K
References (27)
27 references, click to expand
  1. Computed circular dichroism spectra for the evaluation of protein conformation.
    Biochemistry. 1969 Oct;8(10):4108-16 PMID: 5346390
  2. Coassembly of K(V)LQT1 and minK (IsK) proteins to form cardiac I(Ks) potassium channel.
    Nature. 1996 Nov 7;384(6604):80-3 PMID: 8900283
  3. Infrared and laser-Raman spectroscopic studies of thermally-induced globular protein gels.
    Int J Pept Protein Res. 1981 Mar;17(3):353-64 PMID: 7026471
  4. A Fourier transform infrared investigation of the structural differences between ribonuclease A and ribonuclease S.
    Biochim Biophys Acta. 1986 Dec 12;874(3):255-65 PMID: 3790572
  5. Cloning of a membrane protein that induces a slow voltage-gated potassium current.
    Science. 1988 Nov 18;242(4881):1042-5 PMID: 3194754
  6. Aggregation-related conformational change of the membrane-associated coat protein of bacteriophage M13.
    Biochemistry. 1989 Nov 14;28(23):9158-65 PMID: 2690954
  7. Immunohistochemical study of a rat membrane protein which induces a selective potassium permeation: its localization in the apical membrane portion of epithelial cells.
    J Membr Biol. 1990 Jan;113(1):39-47 PMID: 2154581
  8. Estrogen induction of a small, putative K+ channel mRNA in rat uterus.
    Neuron. 1990 May;4(5):807-12 PMID: 2344412
  9. Protein secondary structure of the isolated photosystem II reaction center and conformational changes studied by Fourier transform infrared spectroscopy.
    Biochemistry. 1991 May 7;30(18):4552-9 PMID: 1850626
  10. Beware of proteins in DMSO.
    Biochim Biophys Acta. 1991 Jun 24;1078(2):231-5 PMID: 2065090
  11. The in situ aggregational and conformational state of the major coat protein of bacteriophage M13 in phospholipid bilayers mimicking the inner membrane of host Escherichia coli.
    Biochemistry. 1991 Nov 19;30(46):11147-54 PMID: 1932035
  12. Examination of the peptide sequence requirements for lipid-binding. Alternative pathways for promoting the interaction of amphipathic alpha-helical peptides with phosphatidylcholine.
    Biochim Biophys Acta. 1991 Oct 15;1086(1):106-14 PMID: 1954237
  13. Environment affects amino acid preference for secondary structure.
    Proc Natl Acad Sci U S A. 1992 May 15;89(10):4462-5 PMID: 1584778
  14. Does Fourier-transform infrared spectroscopy provide useful information on protein structures?
    Trends Biochem Sci. 1992 Sep;17(9):328-33 PMID: 1412707
  15. Spectroscopic and functional characterization of the putative transmembrane segment of the minK potassium channel.
    Biochemistry. 1993 Mar 9;32(9):2371-7 PMID: 8443177
  16. Expression of a minimal K+ channel protein in mammalian cells and immunolocalization in guinea pig heart.
    Circ Res. 1993 Nov;73(5):968-73 PMID: 8403266
  17. Secondary structure of M13 coat protein in phospholipids studied by circular dichroism, Raman, and Fourier transform infrared spectroscopy.
    Biochemistry. 1993 Nov 23;32(46):12446-54 PMID: 8241135
  18. Studies of the pore-forming domain of a voltage-gated potassium channel protein.
    Protein Eng. 1994 Feb;7(2):255-62 PMID: 8170928
  19. Changes in activation gating of IsK potassium currents brought about by mutations in the transmembrane sequence.
    FEBS Lett. 1994 Oct 24;353(3):251-4 PMID: 7957868
  20. Integration of a K+ channel-associated peptide in a lipid bilayer: conformation, lipid-protein interactions, and rotational diffusion.
    Biochemistry. 1995 Mar 28;34(12):3893-8 PMID: 7696252
  21. The use and misuse of FTIR spectroscopy in the determination of protein structure.
    Crit Rev Biochem Mol Biol. 1995;30(2):95-120 PMID: 7656562
  22. Effects of trifluoroethanol on the conformations of peptides representing the entire sequence of bovine pancreatic trypsin inhibitor.
    Biochemistry. 1995 Oct 3;34(39):12630-5 PMID: 7548013
  23. Structural characterisation of a slowly activating potassium channel (IsK).
    Biochem Soc Trans. 1995 Aug;23(3):478S PMID: 8566375
  24. MinK residues line a potassium channel pore.
    Neuron. 1996 Mar;16(3):571-7 PMID: 8785054
  25. Cytoplasmic and extracellular IsK peptides activate endogenous K+ and Cl- channels in Xenopus oocytes. Evidence for regulatory function.
    J Biol Chem. 1996 Apr 12;271(15):8768-71 PMID: 8621512
  26. K(V)LQT1 and lsK (minK) proteins associate to form the I(Ks) cardiac potassium current.
    Nature. 1996 Nov 7;384(6604):78-80 PMID: 8900282
  27. Conformational states of a hydrophobic protein. The coat protein of fd bacteriophage.
    Biochemistry. 1978 Apr 4;17(7):1239-46 PMID: 656386
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
Published
1997-07-15
Pages
475-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1218584
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]