NADPH regulates human NAD kinase, a NADP⁺-biosynthetic enzyme.

• Ohashi, Kazuto

• 河井, 重幸

• Koshimizu, Mari

• 村田, 幸作

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(+).

収録刊行物

• Molecular and cellular biochemistry

  Molecular and cellular biochemistry 355 (1-2), 57-64, 2011-09

  Springer Science+Business Media, LLC.

キーワード

• NAD kinase
• Human
• Drug design
• NADP+
• NADPH
• NADH

詳細情報

• CRID: 1050001335720964736
• NII論文ID: 120003405536
• NII書誌ID: AA00745800
• ISSN: 03008177
• HANDLE: 2433/147259
• 本文言語コード: en
• 資料種別: journal article
• データソース種別:
  • IRDB
  • CiNii Articles