Reconstitution of protein targeting to the inner envelope membrane of chloroplasts.

Reconstitution of protein targeting to the inner envelope membrane of chloroplasts.
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DOI:
10.1083/jcb.200605162
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发表时间:
2006-10-23
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Schnell DJ
Schnell DJ
中科院分区:
其他
文献类型:
--
作者:
Li M;Schnell DJ

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叶绿体被膜在光合作用和脂类代谢等关键代谢产物的合成和转运中起着重要作用。尽管这一重要性,包膜膜的生物发生尚未详细研究。为了确定蛋白质靶向内包膜(IM)的决定因素,我们研究了核编码的整合IM蛋白atTic 40的靶向。我们发现,pre-atTic 40被导入叶绿体,并在插入IM之前加工成中间尺寸(int-atTic 40)。Int-atTic 40是可溶的,并从内部基质隔室插入IM中。我们还表明,atTic 40和第二个IM蛋白,atTic 110,可以在体外靶向和插入到分离的IM囊泡。总的来说,我们的实验是一致的“postimport”机制,其中IM蛋白首先从细胞质进口,随后插入到IM从基质。
The chloroplast envelope plays critical roles in the synthesis and regulated transport of key metabolites, including intermediates in photosynthesis and lipid metabolism. Despite this importance, the biogenesis of the envelope membranes has not been investigated in detail. To identify the determinants of protein targeting to the inner envelope membrane (IM), we investigated the targeting of the nucleus-encoded integral IM protein, atTic40. We found that pre-atTic40 is imported into chloroplasts and processed to an intermediate size (int-atTic40) before insertion into the IM. Int-atTic40 is soluble and inserts into the IM from the internal stromal compartment. We also show that atTic40 and a second IM protein, atTic110, can target and insert into isolated IM vesicles in vitro. Collectively, our experiments are consistent with a “postimport” mechanism in which the IM proteins are first imported from the cytoplasm and subsequently inserted into the IM from the stroma.
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