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タイトル: | NADPH regulates human NAD kinase, a NADP⁺-biosynthetic enzyme. |
著者: | Ohashi, Kazuto Kawai, Shigeyuki Koshimizu, Mari Murata, Kousaku |
著者名の別形: | 河井, 重幸 村田, 幸作 |
キーワード: | NAD kinase Human Drug design NADP+ NADPH NADH |
発行日: | Sep-2011 |
出版者: | Springer Science+Business Media, LLC. |
誌名: | Molecular and cellular biochemistry |
巻: | 355 |
号: | 1-2 |
開始ページ: | 57 |
終了ページ: | 64 |
抄録: | NAD kinase (NADK, EC 2.7.1.23) is the sole NADP(+)-biosynthetic enzyme that catalyzes phosphorylation of NAD(+) to yield NADP(+) using ATP as a phosphoryl donor, and thus, plays a vital role in the cell and represents a potentially powerful antimicrobial drug target. Although methods for expression and purification of human NADK have been previously established (Lerner et al. Biochem Biophys Res Commun 288:69-74, 2001), the purification procedure could be significantly improved. In this study, we improved the method for expression and purification of human NADK in Escherichia coli and obtained a purified homogeneous enzyme only through heat treatment and single column chromatography. Using the purified human NADK, we revealed a sigmoidal kinetic behavior toward ATP and the inhibitory effects of NADPH and NADH, but not of NADP(+), on the catalytic activity of the enzyme. These inhibitory effects provide insight into the regulation of intracellular NADPH synthesis. Furthermore, these attributes may provide a clue to design a novel drug against Mycobacterium tuberculosis in which this bacterial NADK is potently inhibited by NADP(+). |
著作権等: | The final publication is available at www.springerlink.com This is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。 |
URI: | http://hdl.handle.net/2433/147259 |
DOI(出版社版): | 10.1007/s11010-011-0838-x |
PubMed ID: | 21526340 |
出現コレクション: | 学術雑誌掲載論文等 |
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