Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase.
Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase.
复制标题
钴腈水合酶的另一种基因类型中发生自身亚基交换
DOI：
10.1371/journal.pone.0050829
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Zhou Z
中科院分区：
文献类型：
--
作者：
Liu Y;Cui W;Xia Y;Cui Y;Kobayashi M;Zhou Z
自身亚基交换是含钴腈水合酶（Co-NHase）家族的翻译后成熟过程之一。所有这些NHase都具有<β-亚基><α-亚基><激活蛋白>的基因组织，这使得激活蛋白在翻译后很容易与NHase的α-亚基形成介导复合物。在这里，我们发现钴掺入另一种具有<α-亚基> <β-亚基> <激活蛋白>基因组织的Co-NHase中，也依赖于自身亚基交换。我们通过在 P14K 基因 N 端添加 Strep 标签，成功分离出恶臭假单胞菌 NRRL-18668 的重组 NHase 激活蛋白 (P14K)。发现 P14K 与 NHase 的 α 亚基形成复合物 [α(StrepP14K)2]。钴掺入恶臭假单胞菌的 NHase 中被证实依赖于含钴 α(StrepP14K)2 和不含钴 NHase 之间的 α 亚基取代。钴被插入到无钴 α(StrepP14K)2 中，但没有插入到无钴 NHase 中，这表明 P14K 不仅充当自身亚基交换伴侣，而且还充当金属伴侣。此外，来自恶臭假单胞菌的NHase也由按<β-亚基><α-亚基><P14K>顺序设计的突变基因表达。我们的研究结果扩展了自亚基交换成熟的一般特征。
Self-subunit swapping is one of the post-translational maturation of the cobalt-containing nitrile hydratase (Co-NHase) family of enzymes. All of these NHases possess a gene organization of <β-subunit> <α-subunit> <activator protein>, which allows the activator protein to easily form a mediatory complex with the α-subunit of the NHase after translation. Here, we discovered that the incorporation of cobalt into another type of Co-NHase, with a gene organization of <α-subunit> <β-subunit> <activator protein>, was also dependent on self-subunit swapping. We successfully isolated a recombinant NHase activator protein (P14K) of Pseudomonas putida NRRL-18668 by adding a Strep-tag N-terminal to the P14K gene. P14K was found to form a complex [α(StrepP14K)2] with the α-subunit of the NHase. The incorporation of cobalt into the NHase of P. putida was confirmed to be dependent on the α-subunit substitution between the cobalt-containing α(StrepP14K)2 and the cobalt-free NHase. Cobalt was inserted into cobalt-free α(StrepP14K)2 but not into cobalt-free NHase, suggesting that P14K functions not only as a self-subunit swapping chaperone but also as a metallochaperone. In addition, NHase from P. putida was also expressed by a mutant gene that was designed with a <β-subunit> <α-subunit> <P14K> order. Our findings expand the general features of self-subunit swapping maturation.
登录
查看更多内容
DOI：
10.1073/pnas.94.22.11986
发表时间：
1997-10-28
影响因子：
11.1
作者：
Kobayashi, M;Fujiwara, Y;Shimizu, S
通讯作者：
Shimizu, S
影响因子：
5.6
作者：
Arakawa, Takatoshi;Kawano, Yoshiaki;Odaka, Masafumi
通讯作者：
Odaka, Masafumi
影响因子：
4.8
作者：
Hashimoto, Y;Hosaka, H;Kobayashi, M
通讯作者：
Kobayashi, M
影响因子：
2.9
作者：
Deng, H;Callender, R;Pei, DH
通讯作者：
Pei, DH
DOI：
10.1016/j.bbrc.2003.10.124
发表时间：
2003-12-12
影响因子：
3.1
作者：
Hourai, S;Miki, M;Yanagi, K
通讯作者：
Yanagi, K