Display Settings:

Items per page
We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 6

2.
Figure 2

Figure 2. Abnormal accumulation of acylcarnitines and triglyceride in the livers of mice lacking SIRT3 during fasting. From: SIRT3 regulates fatty acid oxidation via reversible enzyme deacetylation.

a, b. Metabolomic analyses were conducted on mouse liver tissue (a) and plasma (b), data obtained in SIRT3-/- mice are shown relative to wt mice (n=5/genotype, fasted 24 h); c. Livers extracts from SIRT3-/- and wt mice were analyzed for total phospholipids, triglycerides and cholesterol esters (n=5/genotype, fed or fasted 24 h), *p<0.05

Matthew D. Hirschey, et al. Nature. ;464(7285):121-125.
3.
Figure 5

Figure 5. Mice lacking SIRT3 show reduced ATP production, cold intolerance and hypoglycemia. From: SIRT3 regulates fatty acid oxidation via reversible enzyme deacetylation.

a. Hepatic ATP levels were measured in fed and fasted wt and SIRT3-/- mice (n=5/genotype/condition) b, c. Core temperature (b) and blood glucose (c) were measured in fed and fasted wt and SIRT3-/- mice exposed to cold (4°C) for 6 h (n=5/genotype/condition); *p<0.05, **p<0.01

Matthew D. Hirschey, et al. Nature. ;464(7285):121-125.
4.
Figure 1

Figure 1. Fasting induces SIRT3 expression in oxidative tissues. From: SIRT3 regulates fatty acid oxidation via reversible enzyme deacetylation.

a. Mitochondria were isolated from livers of fed or fasted (6-48 h) wt mice and analyzed for SIRT3 expression by western blot analysis, electron transfer flavoprotein (ETF) was used as a reference; b. Mitochondria isolated from livers of fed or fasted wt and SIRT3-/- mice were analyzed by western blotting analysis with an antiserum specific for anti-acetyllysine, ATP synthase alpha was used as a reference. The arrows identify candidate SIRT3 target proteins: deacetylated during fasting in wt mice, relatively hyperacetylated and not deacetylated during fasting in SIRT3-/- mice.

Matthew D. Hirschey, et al. Nature. ;464(7285):121-125.
5.
Figure 3

Figure 3. Defective fatty acid oxidation in mice lacking SIRT3-/-. From: SIRT3 regulates fatty acid oxidation via reversible enzyme deacetylation.

a, b. Fatty acid oxidation was measured by incubation of liver extract from wt and SIRT3-/- mice with 14C-palmitate, acid-soluble metabolites [(ASM), panel a] and captured CO2 (panel b), n=10/genotype; c. Mitochondrial fatty acid oxidation was measured in other oxidizing tissues, including heart, liver, mixed gastrocnemius and soleus skeletal muscle (SKM), and brown adipose tissue (BAT) (CO2, n=7/genotype, 100 μM substrate), d. Fatty acid oxidation was measured by incubation of 14C-palmitate (100 μM) in liver extract from wt and SIRT3-/- mice one week after injection with adenovirus expression vectors for GFP or SIRT3 (ASM, n=3-4/category); *p<0.05, **p<0.01.

Matthew D. Hirschey, et al. Nature. ;464(7285):121-125.
6.
Figure 4

Figure 4. LCAD is hyperacetylated in SIRT3-/-mice, deacetylated by SIRT3 in vivo and in vitro, and displays increased enzymatic activity when deacetylated. From: SIRT3 regulates fatty acid oxidation via reversible enzyme deacetylation.

a. Liver extracts from wt and SIRT3-/- mice (fed or fasted) were immunoprecipitated with an anti-acetyllysine antiserum and analyzed with anti-LCAD antiserum; b. Expression vectors for wt SIRT3, SIRT3-H248Y (catalytically-inactive SIRT3 mutant), SIRT4, or SIRT5 were co-transfected with expression vectors for FLAG-tagged LCAD and the level of LCAD acetylation was assessed; c. Recombinant LCAD expressed in E. coli was incubated in vitro with recombinant SIRT3 or SIRT3-H248Y, and LCAD acetylation status was assessed; d. Expression vectors for wt SIRT3, SIRT3-H248Y, SIRT4 and SIRT5 (HA-tagged) were co-transfected with expression vectors for FLAG-tagged LCAD and assessed for interaction by co-immunoprecipitation; e. LCAD was expressed and purified with SIRT3 or SIRT3-H248Y and its enzymatic activity measured in vitro using 2, 6 dimethylheptanoyl-CoA as a substrate (n=4 independent assays); f. Recombinant LCAD was expressed in E. coli in the absence (Control) or presence of nicotinamide (NAM, 50 mM), purified and its enzymatic activity measured in vitro using 2, 6 dimethylheptanoyl-CoA as a substrate (n=4 independent assays); g. Expression vectors for wt LCAD, LCAD single acetylation point mutant (LCAD-K42R), or LCAD eight acetylation point mutant (LCAD-8KR) were co-transfected with expression vectors for wt SIRT3 or SIRT3-H248Y, and the level of acetylation was assessed; h. Wild-type LCAD, LCAD-K42R, or LCAD-8KR were expressed, and measured for enzymatic activity in vitro using 2, 6 dimethylheptanoyl-CoA as a substrate (n=5 measurement/sample, error bars represent data two independent protein purifications); *p<0.05, **p<0.01.

Matthew D. Hirschey, et al. Nature. ;464(7285):121-125.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk