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(+).
journal_name
Mol Cell Biochemjournal_title
Molecular and cellular biochemistryauthors
Ohashi K,Kawai S,Koshimizu M,Murata Kdoi
10.1007/s11010-011-0838-xsubject
Has Abstractpub_date
2011-09-01 00:00:00pages
57-64issue
1-2eissn
0300-8177issn
1573-4919journal_volume
355pub_type
杂志文章abstract：:Ehrlich ascites tumor cells incorporate [methyl-3H]thymidine into DNA independently of exogenous growth factors or fetal calf serum. Using an acid/ethanol extraction procedure we have obtained from these tumor cells a fraction that induces both the proliferation and the formation of cell foci by Swiss 3T3 mouse fibrob...
journal_title：Molecular and cellular biochemistry
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更新日期：2004-07-01 00:00:00
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更新日期：2000-09-01 00:00:00
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更新日期：2002-10-01 00:00:00
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doi：10.1007/s11010-017-2954-8
更新日期：2017-06-01 00:00:00
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journal_title：Molecular and cellular biochemistry
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更新日期：1998-11-01 00:00:00
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doi：10.1007/s11010-018-3444-3
更新日期：2019-03-01 00:00:00
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