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. 2000 Mar 20;148(6):1213–1222. doi: 10.1083/jcb.148.6.1213

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Toc64 can be cross-linked to other Toc complex subunits. (a) Outer envelope membranes (40 μg protein) were either not treated or treated with 1 mM CuCl₂ or 1 mM DSP. (b) High molecular weight cross-links form in a concentration-dependent manner in the presence of CuCl₂. Cross-linking was performed for 15 min using the concentrations indicated. Cross-linked products were separated by SDS-PAGE and visualized by staining with Coomassie brilliant blue. Numbers 1 and 3 label identical bands as in a. (c) Prominent cross-linked products (bands 1–4 in a) were cut out of the gel, divided into equal aliquots, and separated by a second SDS-PAGE under reducing conditions. Proteins were analyzed either by silverstaining (left panel) or immunoblotting using the antisera indicated (right panel).