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

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

Content Provider	Semantic Scholar
Author	Ohashi, Kazuto Kawai, Shigeyuki Koshimizu, Mari Murata, Kousaku
Copyright Year	2011
Abstract	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+.
Starting Page	57
Ending Page	64
Page Count	8
File Format	PDF HTM / HTML
DOI	10.1007/s11010-011-0838-x
PubMed reference number	21526340
Journal	Medline
Volume Number	355
Alternate Webpage(s)	https://repository.kulib.kyoto-u.ac.jp/dspace/bitstream/2433/147259/1/s11010-011-0838-x.pdf
Alternate Webpage(s)	https://doi.org/10.1007/s11010-011-0838-x
Journal	Molecular and Cellular Biochemistry
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article

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