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Science 7 April 2000:
Vol. 288. no. 5463, pp. 143 - 146
DOI: 10.1126/science.288.5463.143
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Reports

Unfolding Pathways of Individual Bacteriorhodopsins

F. Oesterhelt, 1 D. Oesterhelt, 2 M. Pfeiffer, 2 A. Engel, 3 H. E. Gaub, 1* D. J. Müller 34

Atomic force microscopy and single-molecule force spectroscopy were combined to image and manipulate purple membrane patches from Halobacterium salinarum. Individual bacteriorhodopsin molecules were first localized and then extracted from the membrane; the remaining vacancies were imaged again. Anchoring forces between 100 and 200 piconewtons for the different helices were found. Upon extraction, the helices were found to unfold. The force spectra revealed the individuality of the unfolding pathways. Helices G and F as well as helices E and D always unfolded pairwise, whereas helices B and C occasionally unfolded one after the other. Experiments with cleaved loops revealed the origin of the individuality: stabilization of helix B by neighboring helices.

1 CeNS and Lehrstuhl für angewandte Physik, Ludwig Maximilians-Universität München, Amalienstrasse 54, 80799 München, Germany.
2 Max-Planck-Institut für Biochemie, Am Klopferspitz 18a, 82152 Martinsried, Germany.
3 M. E. Müller Institute for Structural Biology, Biozentrum, University of Basel, Klingelbergstrasse 70, 4056 Basel, Switzerland.
4 Max-Planck-Insitute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany.
*   To whom correspondence should be addressed.

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