Figure 2.
Figure 2. The ∼42-kD band is a cross-linking product between Tha4 and cpTatC. (A) Thylakoids with radiolabeled cpTatC L231C were incubated with mock translation extract or unlabeled Tha4 P9C followed by mock translation extract or SpF16. After disulfide cross-linking, recovered thylakoids were subjected to denaturing immunoprecipitation with antibody-linked beads as shown above the panels (see Materials and methods). (B) Thylakoids containing radiolabeled cpTatC–His6 L231C were incubated with radiolabeled Tha4 P9C and either SpF12 or mock translation extract and then subjected to disulfide cross-linking. Recovered thylakoids were dissolved in 1% digitonin and cpTatC-His6 purified on magnetic Ni-NTA beads (Materials and methods). Eluted samples in A and B were analyzed by nonreducing or reducing SDS-PAGE/fluorography as in Fig. 1.

The ∼42-kD band is a cross-linking product between Tha4 and cpTatC. (A) Thylakoids with radiolabeled cpTatC L231C were incubated with mock translation extract or unlabeled Tha4 P9C followed by mock translation extract or SpF16. After disulfide cross-linking, recovered thylakoids were subjected to denaturing immunoprecipitation with antibody-linked beads as shown above the panels (see Materials and methods). (B) Thylakoids containing radiolabeled cpTatC–His6 L231C were incubated with radiolabeled Tha4 P9C and either SpF12 or mock translation extract and then subjected to disulfide cross-linking. Recovered thylakoids were dissolved in 1% digitonin and cpTatC-His6 purified on magnetic Ni-NTA beads (Materials and methods). Eluted samples in A and B were analyzed by nonreducing or reducing SDS-PAGE/fluorography as in Fig. 1.

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