Activation of NUDT5, an ADP-ribose pyrophosphatase, by nitric oxide-mediated ADP-ribosylation.
The ADP-ribose (ADPR) pyrophosphatase (ADPRase) NUDT5, a member of a superfamily of Nudix hydrolases, hydrolyzes ADP-ribose (ADPR) to AMP and ribose 5'-phosphate. Nitric oxide (NO) enhances nonenzymatic ADP-ribosylation of proteins such as beta-actin and glyceraldehydes 3-phosphate dehydrogenase in the presence of free ADPR, suggesting a possibility that NUDT5 could also be ADP-ribosylated by its substrate, ADPR. Here, we show that NO stimulates nonenzymatic ADP-ribosylation of NUDT5 using ADP-ribose and consequently activates its ADPRase activity. We found that ADPRase activity in J774 macrophage cells is increased by the treatment with SNP, an exogenous NO generator or TNF-alpha/IFN-gamma, endogenous NO inducers. Anti-NUDT5 antibody pulled down most of the ADPRase activity increased by NO, indicating that the ADPRase regulated by NO is NUDT5. Using recombinant human NUDT5, we also demonstrated that the increase of ADPRase activity is mediated via ADP-ribosylation at cysteine residue(s) in the presence of reductant. This result suggests that NO activates NUDT5 through ADP-ribosylation at cysteine residues of the enzyme in macrophages.
Yu
Hong-Nu
HN
Department of Biochemistry, Institute of Medical Science, Chonbuk National University Medical School, Jeonju, Jeonbuk 560-182, Republic of Korea.
Song
Eun-Kyung
EK
Yoo
Seung-Min
SM
Lee
Young-Rae
YR
Han
Myung-Kwan
MK
Yim
Chang-Yeol
CY
Kwak
Jae-Yong
JY
Kim
Jong-Suk
JS
D000246:Adenosine Diphosphate Ribose / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D003545:Cysteine / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000494:pharmacology*; D011755:Pyrophosphatases / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D014409:Tumor Necrosis Factor-alpha; D000246:Adenosine Diphosphate Ribose; D009569:Nitric Oxide; D007371:Interferon-gamma; C058796:ADP-ribose pyrophosphatase I; C403715:NUDT5 protein, human; D011755:Pyrophosphatases; D003545:Cysteine