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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2019 May 16;39(7):1419–1431. doi: 10.1161/ATVBAHA.118.312346

Figure 7. Peroxynitrite mediates SIRT1 inhibition and zinc release from SIRT1.

Figure 7.

A. SIRT1 inhibition by peroxynitrite (ONOO−). Human recombinant SIRT1 was treated with ONOO− in reaction system for 45 min at 37°C (n=3–6; * P<0.001 vs. control). B. Zinc release from recombinant human SIRT1 with indicated treatments, ONOO− (50 μM) and decomposed ONOO− (dONOO−) (n=3; * P<0.001 vs. ONOO−). Zinc was assayed as described in Methods and was expressed as percentage of maximal zinc release from SIRT1 diluted in 7 M guanidine HCl. C. Exogenous zinc inhibits SIRT1 activity. Human recombinant SIRT1 was treated in advance with nicotinamide (NAM, Catalog # N1788, 400x dilution) or ONOO− (50 μM) with or without ZnCl2 for 15 min, then put in the reaction system for 30 min at 37°C (n=4; * P<0.001 vs. control. # P<0.01 vs. ZnCl2. D. NAD+-binding site or Zn2+-binding module is required for basal SIRT1 activity. Recombinant SIRT1 proteins as indicated above were added in reaction system for 30 min at 37°C. Y, tyrosine; F, phenylalanine; C, cysteine; S, serine. (n=3–4; * P<0.01 vs. WT). E. Scheme of nicotine-induced arterial stiffness via peroxynitrite-mediated SIRT1 inhibition.