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Science 14 March 2003:
Vol. 299. no. 5613, pp. 1747 - 1751
DOI: 10.1126/science.1080945
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Reports

Protein Insertion into the Mitochondrial Inner Membrane by a Twin-Pore Translocase

Peter Rehling,1 Kirstin Model,2 Katrin Brandner,13 Peter Kovermann,4 Albert Sickmann,5 Helmut E. Meyer,5 Werner Kühlbrandt,2 Richard Wagner,4 Kaye N. Truscott,1 Nikolaus Pfanner1*

The mitochondrial inner membrane imports numerous proteins that span it multiple times using the membrane potential Delta psi as the only external energy source. We purified the protein insertion complex (TIM22 complex), a twin-pore translocase that mediated the insertion of precursor proteins in a three-step process. After the precursor is tethered to the translocase without losing energy from the Delta psi , two energy-requiring steps were needed. First, Delta psi acted on the precursor protein and promoted its docking in the translocase complex. Then, Delta psi and an internal signal peptide together induced rapid gating transitions in one pore and closing of the other pore and drove membrane insertion to completion. Thus, protein insertion was driven by the coordinated action of a twin-pore complex in two voltage-dependent steps.

1 Institut für Biochemie und Molekularbiologie, Universität Freiburg, Hermann-Herder-Strasse 7, D-79104 Freiburg, Germany.
2 Max-Planck Institut für Biophysik, Abteilung Strukturbiologie, Heinrich-Hoffmann-Strasse 7, D-60528 Frankfurt am Main, Germany.
3 Fakultät für Biologie, Universität Freiburg, D-79104 Freiburg, Germany.
4 Biophysik, Universität Osnabrück, FB Biologie/Chemie, D-49034 Osnabrück, Germany.
5 Medizinisches Proteom-Center, Gebäude ZKF E/143, Ruhr-Universität Bochum, Universitätsstrasse 150, D-44780 Bochum, Germany.
*   To whom correspondence should be addressed. E-mail: Nikolaus.Pfanner{at}biochemie.uni-freiburg.de

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