Hirohashi T, Hase T, Nakai M
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita 565-0871, Japan.
Plant Physiol. 2001 Apr;125(4):2154-63. doi: 10.1104/pp.125.4.2154.
Preprotein translocation across the outer and inner envelope membranes of chloroplasts is an energy-dependent process requiring ATP hydrolysis. Several precursor proteins analyzed so far have been found to be imported into isolated chloroplasts equally well in the dark in the presence of ATP as in the light where ATP is supplied by photophosphorylation in the chloroplasts themselves. We demonstrate here that precursors of two maize (Zea mays L. cv Golden Cross Bantam) ferredoxin isoproteins, pFdI and pFdIII, show distinct characteristics of import into maize chloroplasts. pFdI, a photosynthetic ferredoxin precursor, was efficiently imported into the stroma of isolated maize chloroplasts both in the light and in the dark. In contrast pFdIII, a non-photosynthetic ferredoxin precursor, was mostly mis-sorted to the intermembrane space of chloroplastic envelopes as an unprocessed precursor form in the light but was efficiently imported into the stroma and processed to its mature form in the dark. The mis-sorted pFdIII, which accumulated in the intermembrane space in the light, could not undergo subsequent import into the stroma in the dark, even in the presence of ATP. However, when the mis-sorted pFdIII was recovered and used for a separate import reaction, pFdIII was capable of import into the chloroplasts in the dark. pFNRII, a ferredoxin-NADP+ reductase isoprotein precursor, showed import characteristics similar to those of pFdIII. Moreover, pFdIII exhibited similar import characteristics with chloroplasts isolated from wheat (Pennisetum americanum) and pea (Pisum sativum cv Alaska). These findings suggest that the translocation of precursor proteins across the envelope membranes of chloroplasts may involve substrate-dependent light-regulated mechanisms.
前体蛋白穿过叶绿体的外膜和内膜是一个依赖能量的过程,需要ATP水解。到目前为止分析的几种前体蛋白已被发现,在黑暗中存在ATP时,它们被导入分离的叶绿体的效率与在光照下相同,在光照下ATP由叶绿体自身的光合磷酸化提供。我们在此证明,两种玉米(Zea mays L. cv Golden Cross Bantam)铁氧还蛋白同工型pFdI和pFdIII的前体,在导入玉米叶绿体时表现出不同的特征。pFdI是一种光合铁氧还蛋白前体,在光照和黑暗条件下都能有效地导入分离的玉米叶绿体基质中。相比之下,pFdIII是一种非光合铁氧还蛋白前体,在光照下大多以未加工的前体形式错误分选到叶绿体被膜的膜间隙中,但在黑暗中能有效地导入基质并加工成成熟形式。在光照下积累在膜间隙中的错误分选的pFdIII,即使在有ATP的情况下,在黑暗中也不能随后导入基质。然而,当回收错误分选的pFdIII并用于单独的导入反应时,pFdIII在黑暗中能够导入叶绿体。铁氧还蛋白-NADP +还原酶同工型前体pFNRII表现出与pFdIII相似的导入特征。此外,pFdIII与从小麦(Pennisetum americanum)和豌豆(Pisum sativum cv Alaska)分离的叶绿体表现出相似的导入特征。这些发现表明,前体蛋白穿过叶绿体被膜的转运可能涉及底物依赖性的光调节机制。