Protein Science, Vol 7, Issue 2 342-348, Copyright © 1998 by Cold Spring Harbor Laboratory Press

ARTICLE

Solid-state NMR studies of the membrane-bound closed state of the colicin E1 channel domain in lipid bilayers

Y. KIM, K. VALENTINE, S. J. OPELLA, S. L. SCHENDEL and W. A. CRAMER
Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104 Current address: LG Chemical Research Park, Daejeon 305-380, Korea.

The colicin E1 channel polypeptide was shown to be organized anisotropically in membranes by solid-state NMR analysis of samples of uniformly (15)N-labeled protein in oriented planar phospholipid bilayers. The 190 residue C-terminal colicin E1 channel domain is the largest polypeptide to have been characterized by (15)N solid-state NMR spectroscopy in oriented membrane bilayers. The (15)N-NMR spectra of the colicin E1 show that: (1) the structure and dynamics are independent of anionic lipid content in both oriented and unoriented samples; (2) assuming the secondary structure of the polypeptide is helical, there are both trans-membrane and in-plane helical segments; (3) trans-membrane helices account for approximately 20-25% of the channel polypeptide, which is equivalent to 38-48 residues of the 190-residue polypeptide. The results of the two-dimensional PISEMA spectrum are interpreted in terms of a single trans-membrane helical hairpin inserted into the bilayer from each channel molecule. These data are also consistent with this helical hairpin being derived from the 38-residue hydrophobic segment near the C-terminus of the colicin E1 channel polypeptide.
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