Evidence for an alpha-helix --> pi-bulge helicity modulation for the neu/erbB-2 membrane-spanning segment. A 1H NMR and circular dichroism study.
The 35-residue peptide corresponding to the very hydrophobic transmembrane region of the tyrosine kinase receptor neu, Neu(TM35), has been synthesized. The peptide can be solubilized in millimolar concentrations in TFE or incorporated into an SDS-water micellar solution or into well-hydrated DMPC/DCPC bicelles. In all these media, circular dichroism demonstrated that the peptide adopts a helical structure for about 80% of its amino acids. The peptide is monomeric below 2 mM in TFE, as also determined by variable concentration experiments. The three-dimensional solution structure in TFE has been obtained by homonuclear proton NMR and shows a well-defined alpha-helix from residues 4 to 21, then a pi-bulge from Ile(22) to Gly(28), and a final short alpha-helix from positions 29 to 32. This experimental finding is in agreement with structures predicted recently by molecular dynamics calculations in a vacuum [Sajot, N., and Genest, M. (2000) Eur. Biophys. J. 28, 648-662]. The biological implications of a possible retention of this structure in a membrane environment are finally discussed.
Goetz
M
M
Institut Européen de Chimie et de Biologie, Ecole Polytechnique, CNRS & Universités de Bordeaux I et II, Talence, France.
Carlotti
C
C
Bontems
F
F
Dufourc
E J
EJ
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008565:Membrane Proteins / Q000138:chemical synthesis / Q000737:chemistry*; D008823:Micelles; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry*; D010742:Phospholipid Ethers / Q000737:chemistry; D017433:Protein Structure, Secondary; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry*; D012967:Sodium Dodecyl Sulfate; D012996:Solutions; D013816:Thermodynamics; D014270:Trifluoroethanol / Q000737:chemistry
D004134:Dimyristoylphosphatidylcholine; D008051:Lipid Bilayers; D008565:Membrane Proteins; D008823:Micelles; D010446:Peptide Fragments; D010742:Phospholipid Ethers; D018719:Erb-b2 Receptor Tyrosine Kinases; D012967:Sodium Dodecyl Sulfate; D012996:Solutions; D014270:Trifluoroethanol; C052298:1,2-dihexadecyl-sn-glycero-3-phosphocholine