Figure 8.

Association of Truncated atTic110N_His and atTic110C_His with Other Tic and Toc Components.

(A) Association of atTic110 deletion constructs with Tic components. Detergent-soluble total chloroplast membranes from wild-type, atTic110_His, atTic110N_His, and atTic110C_His expressing plants were subjected to Ni-NTA chromatography. The Ni-NTA eluates (eluate) were resolved by SDS-PAGE under reducing conditions and immunoblotted with antisera corresponding to the proteins indicated at the left. Lanes 1 to 4 contain 5% of the total membrane fractions (total membrane) used for chromatography. Lanes 9 and 10 are duplicates of lanes 7 and 8 containing threefold more protein.

(B) Association of atTic110 deletion constructs with Toc-Tic supercomplexes. Detergent-soluble total chloroplast membranes from wild-type, atTic110_His, atTic110N_His, and atTic110C_His expressing plants were subjected to immunoaffinity chromatography on anti-atToc33 Sepharose (α-atToc33). The eluates (lanes 4 to 7) were resolved by SDS-PAGE and probed with antibodies corresponding to the proteins indicated at the left of the panels. Lanes 1 to 3 contain 5% of the total membrane fractions (membrane) used for chromatography. The unbound fraction from the atTic110N_His chloroplast extracts that had been immunodepleted on α-atToc33 was subjected to subsequent chromatography on Ni-NTA matrix to precipitate residual atTic110N_His (lane 8).

(C) Association of overexpressed atTic110_His with Toc-Tic supercomplexes. Detergent extracts of total chloroplast membranes from wild-type plants and plants overexpressing atTic110_His were subjected to immunoaffinity chromatography on anti-atToc33 Sepharose (α-atToc33). The eluates (lanes 3 and 4) were resolved by SDS-PAGE and probed with antibodies corresponding to the proteins indicated at the left. Lanes 1 and 2 contain 5% of the total membrane fractions (membrane) used for chromatography.