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Structural Requirements for Membrane Assembly of Proteins Spanning the Membrane Several Times

Lipp, Joachim ; Flint, Nicholas ; Haeuptle, Marie-Theres ; Dobberstein, Bernhard

In: The Journal of Cell Biology, 109 (1989), pp. 2013-2022

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We have investigated the structural requirements for the biogenesis of proteins spanning the membrane several times. Proteins containing various combinations of topological signals (signal anchor and stop transfer sequences) were synthesized in a cell-free translation system and their membrane topology was determined. Proteins spanning the membrane twice were obtained when a signal anchor sequence was followed by either a stop transfer sequence or a second signal anchor sequence. Thus, a signal anchor sequence in the second position can function as a stop transfer sequence, spanning the membrane in the opposite orientation to that of the first signal anchor sequence. A signal anchor sequence in the third position was able to insert amino acid sequences located COOH terminal to it. We conclude that proteins spanning the membrane several times can be generated by stringing together signal anchor and stop transfer sequences. However, not all proteins with three topological signals were found to span the membrane three times. A certain segment located between the first and second topological signal could prevent stable membrane integration of a third signal anchor segment.

Item Type: Article
Journal or Publication Title: The Journal of Cell Biology
Volume: 109
Date Deposited: 15 Jul 2008 17:47
Date: 1989
Page Range: pp. 2013-2022
Faculties / Institutes: Service facilities > Center for Molecular Biology Heidelberg
Subjects: 570 Life sciences
Schriftenreihe ID: Works by Bernhard Dobberstein
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