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The E. coli and Human Nudix hydrolases NudC and NUDT12 cleave damaged NADH
Author(s) -
Ray Atreyei,
Beaupre Brett A.,
Moran Graham R.,
Frick David N.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.606.11
Subject(s) - cleave , biochemistry , enzyme , nad+ kinase , nucleoside , nucleotide , chemistry , biology , gene
Nudix enzymes hydrolyze phosphoanhydride bonds linked to nucleotides. One subclass of Nudix enzymes cleaves dinucleotides to yield two nucleoside monophosphates. The prototype enzyme of this Nudix subclass is the product of the E. coli nudC gene (aka NudC or Orf257), which has been previously shown to cleave NAD(P)H, NAD(P)+, ADP‐ribose, AppA, and NAD‐capped RNA. Since other Nudix enzymes are known to remove potential mutagens from nucleotide pools, we tested the hypothesis that NudC‐like proteins might cleave potentially harmful dinucleotides. Recent studies have shown that many cells express enzymes that cleanse cells of a form of β‐NADH that carries the hydride in the 6‐position of the nicotinamide base (6DHNAD). 6DHNAD is a potent inhibitor of numerous cellular oxidoreductases. In this study, NudC and its human homolog NUDT12 were expressed in E. coli, purified, and their abilities to cleave 6DHNAD were analyzed. The enzymatic activities of each enzyme were quantified using a colorimetric assay and High Pressure Liquid Chromatography. Both proteins required divalent metal ions to hydrolyze 6DHNAD to yield two nucleoside 5′ monophosphates.