Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase.
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 fo...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3511329?pdf=render |
| _version_ | 1819162098084610048 |
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| author | Yi Liu Wenjing Cui Yuanyuan Xia Youtian Cui Michihiko Kobayashi Zhemin Zhou |
| author_facet | Yi Liu Wenjing Cui Yuanyuan Xia Youtian Cui Michihiko Kobayashi Zhemin Zhou |
| author_sort | Yi Liu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T17:22:50Z |
| format | Article |
| id | doaj.art-4ef51af8ef034edf8f79163fbf056e98 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T17:22:50Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-4ef51af8ef034edf8f79163fbf056e982022-12-21T18:18:48ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-01711e5082910.1371/journal.pone.0050829Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase.Yi LiuWenjing CuiYuanyuan XiaYoutian CuiMichihiko KobayashiZhemin ZhouSelf-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.http://europepmc.org/articles/PMC3511329?pdf=render |
| spellingShingle | Yi Liu Wenjing Cui Yuanyuan Xia Youtian Cui Michihiko Kobayashi Zhemin Zhou Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase. PLoS ONE |
| title | Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase. |
| title_full | Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase. |
| title_fullStr | Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase. |
| title_full_unstemmed | Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase. |
| title_short | Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase. |
| title_sort | self subunit swapping occurs in another gene type of cobalt nitrile hydratase |
| url | http://europepmc.org/articles/PMC3511329?pdf=render |
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