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. Author manuscript; available in PMC: 2010 Jun 1.
Published in final edited form as: Plant J. 2009 Feb 3;58(5):831–842. doi: 10.1111/j.1365-313X.2009.03816.x

Figure 5.

Figure 5

In organello competition indicates that cpTatC integration does not proceed via the cpTat or cpSec pathways. Chloroplasts were incubated with 5 mM ATP, 1.5 mM DTT, and either 0.3 M urea (−) or unlabeled bacterially expressed pOE23 or pOE33 (competitor) at the final concentrations depicted above the panels for 7 min in the light. Radiolabeled pcpTatC (a), pOE23 (b), or pOE33 (c) was added to the reaction mixtures and the import reaction allowed to continue for an additional 15 minutes. Chloroplasts from pcpTatC reactions were recovered, lysed, and fractionated into stroma (S) and membranes (M). An aliquot of the membrane fraction was treated with thermolysin (+T). Samples were analyzed by SDS-PAGE and fluorography. Chloroplasts from import reactions with radiolabeled pOE23 or pOE33 were recovered and directly analyzed. Gel band extraction was performed to quantify the relative percentages of stromal and membrane-associated cpTatC, which are represented by numbers below the bands in panel (a). The positions of molecular weight markers (Mr) are shown at left.