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Science 9 August 1996:
Vol. 273. no. 5276, pp. 810 - 812
DOI: 10.1126/science.273.5276.810
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Reports

Organization of Diphtheria Toxin T Domain in Bilayers: A Site-Directed Spin Labeling Study

Kyoung Joon Oh, Hangjun Zhan, * Can Cui, Kálmán Hideg, R. John Collier, dagger Wayne L. Hubbell dagger

The diphtheria toxin transmembrane (T) domain was spin-labeled at consecutive residues in a helical segment, TH9. After binding of the T domain to membranes at low pH, the nitroxide side chains generated by spin labeling were measured with respect to their frequency of collision with polar and nonpolar reagents. The data showed that the helical structure of TH9 in solution is conserved, with one face exposed to water and the other to the hydrophobic interior of the bilayer. Measurement of the depth of the nitroxide side chains from the membrane surfaces revealed an incremental change of about 5 angstroms per turn, which is consistent with a transmembrane orientation of an alpha  helix. These results indicate that the helix forms the lining of a transmembrane water-filled channel.

K. J. Oh and W. L. Hubbell, Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095-7008, USA.
H. Zhan, C. Cui, R. J. Collier, Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.
K. Hideg, Central Research Laboratory, Chemistry, University of Pécs, H-7643 Pécs, Hungary.
*   Present address: Arris Pharmaceutical Corporation, 385 Oyster Point Boulevard, Suite 3, South San Francisco, CA 94080, USA.

dagger    To whom correspondence should be addressed.


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