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. 2010 Mar 25;29(9):1477–1488. doi: 10.1038/emboj.2010.44

Figure 7.

Figure 7

Cross-linked precursor dimer can be transported without breaking the linkage. tOE17-20F3C precursor was incubated with thylakoids in binding reactions without any additions (lanes 1, 2, 7 and 8) or with 0.5 mM TCEP to prevent disulphide formation (lanes 3–6, 9–12). The thylakoids were then subjected to cross-linking with BMOE from a DMSO stock solution (lanes 5, 6, 11 and 12), mock treated with DMSO (lanes 3, 4, 9 and 10) or mock treated with import buffer (IB). The thylakoids were buffer washed without (lanes 1, 2, 7 and 8) or with (lanes 3–6, 9–12) 2 mM DTT. Recovered thylakoids were either analysed directly as binding samples (lane B) or used for further chase assays as described in Materials and methods (lane C). Aliquots of the recovered thylakoids were treated with the thermolysin (lanes 7–12). Samples were analysed by non-reducing SDS–PAGE and fluorography (A) or reducing SDS–PAGE and fluorography (B).