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  • Chloroplast Outer Membrane β-Barrel Proteins Use Components of the General Import Apparatus.

Chloroplast Outer Membrane β-Barrel Proteins Use Components of the General Import Apparatus.

The Plant cell (2019-06-21)
Philip M Day, Kentaro Inoue, Steven M Theg
摘要

Chloroplasts evolved from a cyanobacterial endosymbiont that resided within a eukaryotic cell. Due to their prokaryotic heritage, chloroplast outer membranes contain transmembrane β-barrel proteins. While most chloroplast proteins use N-terminal transit peptides to enter the chloroplasts through the translocons at the outer and inner chloroplast envelope membranes (TOC/TIC), only one β-barrel protein, Toc75, has been shown to use this pathway. The route other β-barrel proteins use has remained unresolved. Here we use in vitro pea (Pisum sativum) chloroplast import assays and transient expression in Nicotiana benthamiana to address this. We show that a paralog of Toc75, outer envelope protein 80 kD (OEP80), also uses a transit peptide but has a distinct envelope sorting signal. Our results additionally indicate that β-barrels that do not use transit peptides also enter the chloroplast using components of the general import pathway.

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Sigma-Aldrich
胰蛋白酶 来源于牛胰腺, TPCK Treated, essentially salt-free, lyophilized powder, ≥10,000 BAEE units/mg protein
Sigma-Aldrich
Protease Inhibitor Cocktail, for plant cell and tissue extracts, DMSO solution
Sigma-Aldrich
胰蛋白酶抑制剂 来源于大豆, lyophilized powder
Sigma-Aldrich
嗜热菌蛋白酶 from Geobacillus stearothermophilus, Type X, lyophilized powder, 30-350 units/mg protein (E1%/280)
Sigma-Aldrich
Anti-T7 Epitope Tag Antibody, T7 (MASMTGGQQMG), Chemicon®, from rabbit