Formation of irregular giant peroxisomes by overproduction of the crystalloid core protein methanol oxidase in the methylotrophic yeast Hansenula polymorpha
Formation of irregular giant peroxisomes by overproduction of the crystalloid core protein methanol oxidase in the methylotrophic yeast Hansenula polymorpha
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甲基营养酵母多形汉逊酵母中晶体核心蛋白甲醇氧化酶的过量产生形成不规则的巨型过氧化物酶体
DOI：
--
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发表时间：
1989
影响因子：
5.3
通讯作者：
K. Kowallik
中科院分区：
文献类型：
--
作者：
R. Roggenkamp;T. Didion;K. Kowallik
甲基营养酵母汉逊酵母的过氧化物酶体中的晶体核心由八聚黄素蛋白甲醇氧化酶 (MOX) 组成。我们用含有克隆 MOX 基因的高拷贝数载体转化酵母细胞，以研究其对调节、蛋白质输入和过氧化物酶体生物合成的影响。在转化的野生型细胞中，未检测到 MOX 表达的增加。通过特定的选择程序分离出 MOX 活性缺陷的突变体。描述了两种结构的 MOX 突变体，它们允许在转化时过量产生完全活性的酶，其数量约为总细胞蛋白的三分之二。过量产生的蛋白质被输入到过氧化物酶体中，改变了它们的形态（在薄片中）和细胞裂解物中的稳定性；细胞器呈现出形成矩形体的倾向，并且其管腔完全被晶体结构填充。与未转化酵母细胞的大小相比，过氧化物酶体的总体大小增加了数倍。结果表明，单一蛋白质的过量生产和输入赋予了过氧化物酶体高生长能力。
The crystalloid core in peroxisomes of the methylotrophic yeast Hansenula polymorpha is composed of the octameric flavoprotein methanol oxidase (MOX). We transformed yeast cells with a high-copy-number vector harboring the cloned MOX gene in order to study the effects on regulation, protein import, and peroxisome biosynthesis. In transformed wild-type cells, no increase in expression of MOX was detectable. Mutants defective in MOX activity were isolated by a specific selection procedure. Two structural MOX mutants are described that allow overproduction of a fully active enzyme upon transformation at quantities of about two-thirds of the total cellular protein. The overproduced protein was imported into peroxisomes, altering their morphology (in thin sections) and stability in cell lysates; the organelles showed a tendency to form rectangular bodies, and their lumina were completely filled with the crystalloid structure. The overall size of the peroxisomes was increased severalfold in comparison with the size of nontransformed yeast cells. The results suggest high capacities of peroxisomal growth conferred by overproduction and import of a single protein.
二羟基丙酮合酶是博伊丁假丝酵母甲醇诱导的过氧化物酶体的丰富成分。
DOI：
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发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Goodman,JM
通讯作者：
Goodman,JM
DOI：
10.1016/s0021-9258(17)39756-9
发表时间：
1984-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J M Goodman;C W Scott;P N Donahue;J P Atherton
通讯作者：
J M Goodman;C W Scott;P N Donahue;J P Atherton