Product Details
| Alternative Name: | Sirtuin 5, Sir2-like protein 5, SIR2L5, NAD-dependent deacetylase Sirtuin-5 |
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| MW: | ~32.3kDa |
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| Source: | Produced in E. coli. |
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| UniProt ID: | Q9NXA8 |
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| Formulation: | Liquid. In 25mM TRIS, pH 7.5, 100mM sodium chloride, 5 mM DTT, containing 10% (v/v) glycerol. |
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| Purity: | ≥95% (SDS-PAGE) |
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| Purity Detail: | Purified by multi-step chromatography. |
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| Specific Activity: | One unit = 1 pmol/min at 37°C, 250µM FLUOR DE LYS® Succinyl, Desuccinylase Substrate (Prod. No. BML-KI590), 2mM NAD+. |
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| Application Notes: | Useful for studies of enzyme kinetics, modulator activity, drug discovery. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Use/Stability: | The enzyme is stable on ice for the time typically required to set up an experiment (30-60 min.), but may lose activity with prolonged storage on ice. It is recommended that thawing and dilution of the enzyme be done within as short a time as possible before start of the assay. The remaining, unused enzyme should be refrozen quickly by, for example, snap freezing in a dry/ice ethanol bath or liquid nitrogen. The enzyme is stable to at least 4 freeze/thaw cycles. To minimize the number of freeze/thaw cycles, aliquot the SIRT5 into separate tubes and store at -80° C.NOTE: When stored under the above conditions, this enzyme is stable at the concentration supplied, in its current storage buffer. Procedures such as dilution of the enzyme followed by refreezing, could lead to loss of activity. |
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| Handling: | Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -80°C. |
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| Scientific Background: | SIRT5, along with SIRTs 3 and 4, is one of three human mitochondrial sirtuins (homologs of yeast Sir2). SIRT5, which is localized to both the mitochondrial matrix and the intermembrane space, is an NAD+-dependent lysine deacetylase (class III HDAC). SIRT5's only known physiological target is the intermembrane space protein, cytochrome c, suggesting the possibility of regulatory roles in metabolism and/or apoptosis. SIRT5 has also been shown to deacetylate and thereby upregulate the matrix urea cycle enzyme carbamoyl phosphate synthetase 1. This implies an important role for SIRT5 in ammonia detoxification and adaptation to elevated amino acid catabolism during starvation, calorie restriction or a high protein diet. SIRT5 is most strongly expressed in thymus, lymphoblasts and heart muscle cells and its chromosomal location (6p23) has been implicated in chromosomal abnormalities associated with malignancies. SIRT5 deacetylase activity may be measured with the FLUOR DE LYS® Desuccinylase (Prod. No. BML-KI590) or SIRT1 (Prod. No. BML-KI177) substrates, and, like SIRT1, its activity is stimulated by polyphenols such as resveratrol and fisetin. Two SIRT5 crystal structures have been determined, one as a complex with ADP-ribose and the other a complex with the inhibitor suramin. This recombinant preparation, expressed in E. coli, comprises residues 37-310 of SIRT5 isoform 1, with an N-terminal His-tag. Recombinant human mitochondrial processing protease cleaves full-length SIRT5 after residue thirty-six, suggesting that the mature, in vivo form may begin with residue 37. Consistent with this, N-terminal sequence of mouse SIRT5 processed in 293T cells also commences at residue 37. |
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| Regulatory Status: | RUO - Research Use Only |
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SDS-PAGE Analysis: Lane 1: MWM; Lane 2: 1.0 µg of purified SIRT5 protein.
Figure 1
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Product Literature References
Sirtuin 1 Inhibiting Thiocyanates (S1th)—A New Class of Isotype Selective Inhibitors of NAD+ Dependent Lysine Deacetylases: N. Wossner, et al.; Front. Oncol.
10, 657 (2020),
Abstract;
Full Text
Identification of Diketopiperazine-Containing 2-Anilinobenzamides as Potent Sirtuin 2 (SIRT2)-Selective Inhibitors Targeting the "Selectivity Pocket", Substrate-Binding Site, and NAD+-Binding Site: P. Mellini, et al.; J. Med. Chem.
62, 5844 (2019),
Abstract;
Pharmacophore modeling and virtual screening studies to identify novel selective SIRT2 inhibitors: G. Eren, et al.; J. Mol. Graph. Model.
10, 1313 (2019),
Abstract;
SIRT4 Is a Lysine Deacylase that Controls Leucine Metabolism and Insulin Secretion: K.A. Anderson, et al.; Cell Metab.
25, 838 (2017),
Abstract;
Full Text
Sirtuins regulate proteomic responses near thermal tolerance limits in the blue mussels Mytilusgalloprovincialis and Mytilus trossulus: M.C. Vasquez, et al.; J. Exp. Biol.
220, 4515 (2017),
Abstract;
Investigating the Sensitivity of NAD+-Dependent Sirtuin Deacylation Activities to NADH: A.S. Madsen, et al.; J. Biol. Chem.
291, 7128 (2016),
Application(s): Incubation, measurement of hydrolase activity,
Abstract;
Role of deleted in breast cancer 1 (DBC1) protein in SIRT1 deacetylase activation induced by protein kinase A and AMP-activated protein kinase: V. Nin, et al.; J. Biol. Chem.
287, 23489 (2012),
Abstract;
Full Text
General Literature References
SIRT5 Deacetylates carbamoyl phosphate synthetase 1 and regulates the urea cycle: T. Nakagawa, et al.; Cell
137, 560 (2009),
Abstract;
Opposing effects of sirtuins on neuronal survival: SIRT1-mediated neuroprotection is independent of its deacetylase activity: J.A. Pfister, et al.; PLoS ONE
3, e4090 (2008),
Abstract;
Substrates and regulation mechanisms for the human mitochondrial sirtuins Sirt3 and Sirt5: C. Schlicker, et al.; J. Mol. Biol.
382, 790 (2008),
Abstract;
Structural basis of inhibition of the human NAD+-dependent deacetylase SIRT5 by suramin: A. Schuetz, et al.; Structure
15, 377 (2007),
Abstract;
Assignment of the NAD-dependent deacetylase sirtuin 5 gene (SIRT5) to human chromosome band 6p23 by in situ hybridization: U. Mahlknecht, et al.; Cytogenet. Genome Res.
112, 208 (2006),
Abstract;
Compositions and methods for selectively activating human sirtuins: K.T. Howitz & R.E. Zipkin; U.S. Patent Application 20060014705, (2006),
Evolutionarily conserved and nonconserved cellular localizations and functions of human SIRT proteins: E. Michishita, et al.; Mol. Biol. Cell
16, 4623 (2005),
Abstract;
The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase:: B.J. North, et al.; Mol. Cell
11, 437 (2003),
Abstract;
Characterization of five human cDNAs with homology to the yeast SIR2 gene: Sir2-like proteins (sirtuins) metabolize NAD and may have protein ADP-ribosyltransferase activity: R.A. Frye; Biochem. Biophys. Res. Commun.
260, 273 (1999),
Abstract;
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