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ツゲ ヒデアキ
TSUGE HIDEAKI
津下 英明 所属 京都産業大学 生命科学部 先端生命科学科 職種 教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 2005/08 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | A second novel dye-linked L-proline dehydrogenase complex is present in the hyperthermophilic archaeon Pyrococcus horikoshii OT-3 |
| 執筆形態 | その他 |
| 掲載誌名 | FEBS JOURNAL |
| 出版社・発行元 | WILEY-BLACKWELL |
| 巻・号・頁 | 272(16),pp.4044-4054 |
| 著者・共著者 | R Kawakami,H Sakuraba,H Tsuge,S Goda,N Katunuma,T Ohshima |
| 概要 | Two distinguishable activity bands for dye-linked L-proline dehydrogenase (PDH1 and PDH2) were detected when crude extract of the hyperthermophilic archaeon Pyrococcus horikoshii OT-3 was run on a polyacrylamide gel. After purification, PDH1 was found to be composed of two different subunits with molecular masses of 56 and 43 kDa, whereas PDH2 was composed of four different subunits with molecular masses of 52, 46, 20 and 8 kDa. The native molecular masses of PDH1 and PDH2 were 440 and 101 kDa, respectively, indicating that PDH1 has an alpha(4)beta(4) structure, while PDH2 has an alpha(4)beta(4) structure. PDH2 was found to be similar to the dye-linked L-proline dehydrogenase complex from Thermococcus profundus, but PDH1 is a different type of enzyme. After production of the enzyme in Escherichia coli, high-performance liquid chromatography showed the PDH1 complex to contain the flavins FMN and FAD as well as ATP. Gene expression and biochemical analyses of each subunit revealed that the beta subunit bound FAD and exhibited proline dehydrogenase activity, while the alpha subunit bound ATP, but unlike the corresponding subunit in the T. profundus enzyme, it exhibited neither proline dehydrogenase nor NADH dehydrogenase activity. FMN was not bound to either subunit, suggesting it is situated at the interface between the alpha and beta subunits. A comparison of the amino-acid sequences showed that the ADP-binding motif in the alpha subunit of PDH1 clearly differs from that in the alpha subunit of PDH2. It thus appears that a second novel dye-linked L-proline dehydrogenase complex is produced in P. horikoshii. |
| DOI | 10.1111/j.1742-4658.2005.04810.x |
| ISSN | 1742-464X/1742-4658 |