Lipid modifications of a ras peptide exhibit altered packing and mobility versus host membrane as detected by2H solid-state NMR

Alexander Vogel, Catherine P. Katzka, Herbert Waldmann, Klaus Arnold, Michael F Brown, Daniel Huster

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

The human N-ras protein binds to cellular membranes by insertion of two covalently bound posttranslational lipid modifications, which is crucial for its function in signal transduction and cell proliferation. Mutations in ras may lead to unregulated cell growth and eventually cancer, making it an important therapeutic target. Here we have investigated the molecular details of the membrane binding mechanism. A heptapeptide derived from the C-terminus of the human N-ras protein was synthesized including two hexadecyl modifications. Solid-state 2H NMR was used to determine the packing and molecular dynamics of the ras lipid chains as well as the phospholipid matrix. Separately labeling the chains of the peptide and the phospholipids with 2H enabled us to obtain atomically resolved parameters relevant to their structural dynamics. While the presence of ras only marginally affected the packing of DMPC membranes, dramatically lower order parameters (SCD) were observed for the ras acyl chains indicating modified packing properties. Essentially identical projected lengths of the 16:0 ras chains and the 14:0 DMPC chains were found, implying that the polypeptide backbone is located at the lipid-water interface. Dynamical properties of both the ras and phospholipid chains were determined from spin-lattice 2H relaxation (R 1Z) measurements. Plots of R1Z rates versus the corresponding squared segmental order parameters revealed striking differences. We propose the ras peptide is confined to microdomains containing DMPC chains which are in exchange with the bulk bilayer on the 2H NMR time scale (∼10-5 s). Compared to the host DMPC matrix, the ras lipid modifications are extremely flexible and undergo relatively large amplitude motions. It is hypothesized that this flexibility is a requirement for the optimal anchoring of lipid-modified proteins to cellular membranes.

Original languageEnglish (US)
Pages (from-to)12263-12272
Number of pages10
JournalJournal of the American Chemical Society
Volume127
Issue number35
DOIs
StatePublished - Sep 7 2005

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Dimyristoylphosphatidylcholine
Lipids
Peptides
Nuclear magnetic resonance
Membranes
Phospholipids
ras Proteins
Proteins
Signal transduction
Spin-lattice relaxation
Polypeptides
Structural dynamics
Cell proliferation
Cell growth
Molecular Dynamics Simulation
Post Translational Protein Processing
Labeling
Molecular dynamics
Signal Transduction
Cell Proliferation

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Lipid modifications of a ras peptide exhibit altered packing and mobility versus host membrane as detected by2H solid-state NMR. / Vogel, Alexander; Katzka, Catherine P.; Waldmann, Herbert; Arnold, Klaus; Brown, Michael F; Huster, Daniel.

In: Journal of the American Chemical Society, Vol. 127, No. 35, 07.09.2005, p. 12263-12272.

Research output: Contribution to journalArticle

Vogel, Alexander ; Katzka, Catherine P. ; Waldmann, Herbert ; Arnold, Klaus ; Brown, Michael F ; Huster, Daniel. / Lipid modifications of a ras peptide exhibit altered packing and mobility versus host membrane as detected by2H solid-state NMR. In: Journal of the American Chemical Society. 2005 ; Vol. 127, No. 35. pp. 12263-12272.
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