Dual role of Zn2+ in maintaining structural integrity and suppressing deacetylase activity of SIRT1.
Zn(2+) directly participates in catalysis of histone deacetylase (HDAC) Classes I, II, IV enzymes while its role in HDAC Class III activity is not well established. Herein we investigated the effects of Zn(2+) on the deacetylase activity of sirtuin 1 (silent mating type information regulation 2 homolog 1, SIRT1). We found that the inherent Zn(2+) at the zinc-finger motif of SIRT1 is essential for the structural integrity and the deacetylase activity of SIRT1, whereas the exogenous Zn(2+) strongly inhibits the deacetylase activity with an IC(50) of 0.82muM for Zn(Gly)(2). SIRT1 activity suppressed by the exogenous Zn(2+) can be fully recovered by the metal chelator EDTA but not by the activator resveratrol. We also identified Zn(2+) as a noncompetitive inhibitor for the substrates of NAD(+) and the acetyl peptide P53-AMC. The 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence titration experiments and site-directed mutagenesis study suggested that the exogenous Zn(2+) binds to SIRT1 but not at the zinc-finger motif. These results indicate that Zn(2+) plays a dual role in SIRT1 activity. Inherent Zn(2+) at the zinc-finger motif is structurally related and essential for SIRT1 activity. On the other hand, Zn(2+) may also bind to another site different from the zinc-finger motif or the binding sites for the substrates or resveratrol and act as a potent inhibitor of SIRT1.
Chen
Lei
L
Department of Molecular Pharmacology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu-Chong-Zhi Road, Shanghai 201203, China.
Feng
Yu
Y
Zhou
Yinqiu
Y
Zhu
Weiliang
W
Shen
Xu
X
Chen
Kaixian
K
Jiang
Hualiang
H
Liu
Dongxiang
D
D000107:Acetylation / Q000187:drug effects; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D001667:Binding, Competitive; D002384:Catalysis / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D009154:Mutation; D009243:NAD / Q000378:metabolism; D011485:Protein Binding; D000077185:Resveratrol; D056564:Sirtuin 1 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013267:Stilbenes / Q000494:pharmacology; D013379:Substrate Specificity; D015032:Zinc / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D016335:Zinc Fingers / Q000235:genetics
D002614:Chelating Agents; D013267:Stilbenes; D009243:NAD; D004492:Edetic Acid; C447939:SIRT1 protein, human; D056564:Sirtuin 1; D015032:Zinc; D000077185:Resveratrol