Bowler C, Alliotte T, Van den Bulcke M, Bauw G, Vandekerckhove J, Van Montagu M, Inzé D
Laboratorium voor Genetica, Rijksuniversiteit Gent, Belgium.
Proc Natl Acad Sci U S A. 1989 May;86(9):3237-41. doi: 10.1073/pnas.86.9.3237.
In the plant Nicotiana plumbaginifolia, manganese superoxide dismutase (MnSOD) is synthesized in the cytoplasm as a preprotein and is subsequently translocated to the mitochondrial matrix with corresponding cleavage of an NH2-terminal leader sequence. To determine whether the plant enzyme could replace the endogenous SOD activities of Escherichia coli and yeast, constructions have been made in appropriate vectors for expression of the preprotein and the mature MnSOD. These were introduced into SOD-deficient strains for complementation studies. In E. coli, both forms of the protein were shown to be active and able to complement SOD deficiency to different degrees. Expression of the preprotein in a yeast strain lacking a mitochondrial MnSOD resulted in a restoration of wild-type growth, only possible if the plant protein was being targeted to the mitochondria. Subsequent studies revealed that the protein was processed and that the leader sequence was cleaved at the identical position as recognized by the mitochondrial peptidase of plants. The components mediating mitochondrial import thus appear to be highly conserved between plants and yeast.
在植物蓝茉莉叶烟草中,锰超氧化物歧化酶(MnSOD)最初作为前体蛋白在细胞质中合成,随后伴随着NH2末端前导序列的相应切割而转运至线粒体基质。为了确定这种植物酶是否能够替代大肠杆菌和酵母的内源性超氧化物歧化酶活性,已构建了合适的载体用于前体蛋白和成熟MnSOD的表达。将这些载体导入超氧化物歧化酶缺陷型菌株中进行互补研究。在大肠杆菌中,两种形式的蛋白均显示具有活性,并且能够不同程度地互补超氧化物歧化酶缺陷。在缺乏线粒体MnSOD的酵母菌株中表达前体蛋白,导致野生型生长得以恢复,前提是植物蛋白能够靶向至线粒体。后续研究表明该蛋白被加工处理,并且前导序列在与植物线粒体肽酶识别的相同位置被切割。因此,介导线粒体导入的成分在植物和酵母之间似乎高度保守。