TIC62 redox-regulated translocon composition and dynamics.
The preprotein translocon at the inner envelope of chloroplasts (Tic complex) facilitates the import of nuclear-encoded preproteins into the organelle. Seven distinct subunits have been identified so far. For each of those, specific functions have been proposed based on structural prediction or experimental evidence. Three of those subunits possess modules that could act as redox-active regulatory components in the import process. To date, however, the mode of redox regulation of the import process remains enigmatic. To investigate how the chloroplast redox state influences translocon behavior and composition, we studied the Tic component and the putative redox sensor Tic62 in more detail. The experimental results provide evidence that Tic62 can act as a bona fide dehydrogenase in vitro, and that it changes its localization in the chloroplast dependent on the NADP+/NADPH ratio in the stroma. Moreover, the redox state influences the interactions of Tic62 with the translocon and the flavoenzyme ferredoxin-NADP+ oxidoreductase. Additionally, we give initial experimental insights into the Tic62 structure using circular dichroism measurements and demonstrate that the protein consists of two structurally different domains. Our results indicate that Tic62 possesses redox-dependent properties that would allow it to fulfill a role as redox sensor protein in the chloroplast.
Stengel
Anna
A
Munich Center for Integrated Protein Science CiPS, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strasse 25, D-81377 Munich, Germany.
Benz
Philipp
P
Balsera
Mónica
M
Soll
Jürgen
J
Bölter
Bettina
B
D000595:Amino Acid Sequence; D002736:Chloroplasts / Q000378:metabolism; D002942:Circular Dichroism; D005288:Ferredoxins / Q000737:chemistry; D018506:Gene Expression Regulation, Plant; D008954:Models, Biological; D008969:Molecular Sequence Data; D009249:NADP / Q000737:chemistry*; D009252:NADPH Dehydrogenase / Q000378:metabolism* / Q000502:physiology; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D018532:Pisum sativum / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D017386:Sequence Homology, Amino Acid
D005288:Ferredoxins; D009249:NADP; D010088:Oxidoreductases; D009252:NADPH Dehydrogenase; D010100:Oxygen