Article
Received: 8 September 2008; Revised: 5 December 2008; Accepted: 8 December 2008
10.1002/pro.50 About DOI
Biochemical characterization, localization, and tissue distribution of the longer form of mouse SIRT3 |
| Lei Jin 1 *, Heidi Galonek 1, Kristine Israelian 1, Wendy Choy 1, Michael Morrison 1, Yu Xia 2 a, Xiaohong Wang 2 a, Yihua Xu 2 a, Yuecheng Yang 2 a, Jesse J. Smith 1, Ethan Hoffmann 1, David P. Carney 1, Robert B. Perni 1, Michael R. Jirousek 1, Jean E. Bemis 1, Jill C. Milne 1, David A. Sinclair 1, Christoph H. Westphal 1 |
| 1Sirtris, a GSK Company, 200 Technology Square, Cambridge, MA 02139 2Shanghai Medicilon Inc., 67 Libing Road, Building 5, Zhangjiang High-Tech Park, Shanghai 201203, China |
| email: Lei Jin (ljin@sirtrispharma.com) |
*Correspondence to Lei Jin, Sirtris, a GSK Company, 200 Technology Square, Cambridge, MA 02139
aCurrent address: Viva Biotech Ltd, 1043 Halei Road, Suite 502, Zhangjiang High-Tech Park, Shanghai 201203, China
aCurrent address: Viva Biotech Ltd, 1043 Halei Road, Suite 502, Zhangjiang High-Tech Park, Shanghai 201203, China
aCurrent address: Viva Biotech Ltd, 1043 Halei Road, Suite 502, Zhangjiang High-Tech Park, Shanghai 201203, China
aCurrent address: Viva Biotech Ltd, 1043 Halei Road, Suite 502, Zhangjiang High-Tech Park, Shanghai 201203, China
| Keywords |
| sirtuin NAD+-dependent deacetylation inhibitor mitochondrial function |
| Abstract |
SIRT3 is a key mitochondrial protein deacetylase proposed to play key roles in regulating mitochondrial metabolism but there has been considerable debate about its actual size, the sequences required for activity, and its subcellular localization. A previously cloned mouse SIRT3 has high sequence similarity with the C-terminus of human SIRT3 but lacks an N-terminal mitochondrial targeting sequence and has no detectable deacetylation activity in vitro. Using 5 rapid amplification of cDNA ends, we cloned the entire sequence of mouse SIRT3, as well as rat and rabbit SIRT3. Importantly, we find that full-length SIRT3 protein localizes exclusively to the mitochondria, in contrast to reports of SIRT3 localization to the nucleus. We demonstrate that SIRT3 has no deacetylation activity in vitro unless the protein is truncated, consistent with human SIRT3. In addition, we determined the inhibition constants and mechanism of action for nicotinamide and a small molecule SIRT3 inhibitor against active mouse SIRT3 and show that the mechanisms are different for the two compounds with respect to peptide substrate and NAD+. Thus, identification and characterization of the actual SIRT3 sequence should help resolve the debate about the nature of mouse SIRT3 and identify new mechanisms to modulate enzymatic activity. |
Received: 8 September 2008; Revised: 5 December 2008; Accepted: 8 December 2008
| Digital Object Identifier (DOI) |
10.1002/pro.50 About DOI
