Regulation of two GTPases Toc159 and Toc34 in the translocon of the outer envelope of chloroplasts

Abstract

The GTPases Toc159 and Toc34 of the translocon of the outer envelope of chloroplasts (TOC) are involved in recognition and transfer of precursor proteins at the cytosolic face of the organelle. Both proteins engage multiple interactions within the translocon during the translocation process, including dimeric states of their G-domains. The units of the Toc34 homodimer are involved in the recognition of the transit peptide representing the translocation signal of precursor proteins. This substrate recognition is part of the regulation of the GTPase cycle of Toc34. The Toc159 monomer and the Toc34 homodimer recognize the transit peptide of the small subunit of Rubisco at the N- and at the C-terminal region, respectively. Analysis of the transit peptide interaction by crosslinking shows that the heterodimer between both G-domains binds pSSU most efficiently. While substrate recognition by Toc34 homodimer was shown to regulate nucleotide exchange, we provide evidence that the high activation energy of the GTPase Toc159 is lowered by substrate recognition. The nucleotide affinity of Toc34G homodimer and Toc159G monomer are distinct, Toc34G homodimer recognizes GDP and Toc159G GTP with highest affinity. Moreover, the analysis of the nucleotide association rates of the monomeric and dimeric receptor units suggests that the heterodimer has an arrangement distinct from the homodimer of Toc34. Based on the biochemical parameters determined we propose a model for the order of events at the cytosolic side of TOC. The molecular processes described by this hypothesis range from transit peptide recognition to perception of the substrate by the translocation channel. mehr

Mehr zum Titel

Titel	Regulation of two GTPases Toc159 and Toc34 in the translocon of the outer envelope of chloroplasts
Medien	Biochimica Et Biophysica Acta. Proteins and Proteomics
Heft	6
Band	1867
ISSN	1878-1454
Verfasser	Katharina Wiesemann, Prof. Dr. Stefan Simm, Oliver Mirus, Roman Ladig, Enrico Schleiff
Seiten	627–636
Veröffentlichungsdatum	6.2019
Zitation	Wiesemann, Katharina; Simm, Stefan; Mirus, Oliver; Ladig, Roman; Schleiff, Enrico (2019): Regulation of two GTPases Toc159 and Toc34 in the translocon of the outer envelope of chloroplasts. Biochimica Et Biophysica Acta. Proteins and Proteomics 1867 (6), 627–636. DOI: 10.1016/j.bbapap.2019.01.002