Figure 6.
The FtsH2 mature domain is incompatible for transport by the Sec-pathway. Radiolabeled TP2Mat5 (a) or TP5Mat2 (b) as diagrammed below the panels was incubated with intact pea chloroplasts in a reaction containing 5 mM ATP and light at 25 °C for 20 min, in the presence or absence of nigericin and valinomycin (N/V) or sodium azide (SA). Intact chloroplasts were recovered from reaction and treated with thermolysin (C+). Untreated intact chloroplasts were fractionated into stroma (S) and thylakoids. Thylakoid aliquots were washed with import buffer (T) or treated with thermolysin (T+). Samples were analyzed by SDS-PAGE and fluorography on 7.5% gels except for the T+ samples, which were analyzed on a 12.5% gel. Arrows on diagrams denote swapping points between FtsH5 and FtsH2 sequences.