scholarly journals Mitochondria-targeted inhibitors of the human SIRT3 lysine deacetylase

2021 ◽  
Author(s):  
Kathrin S. Troelsen ◽  
Michael Bæk ◽  
Alexander L. Nielsen ◽  
Andreas S. Madsen ◽  
Nima Rajabi ◽  
...  
Keyword(s):  
Selective Inhibition ◽  
Mitochondrial Proteins ◽  
Lysine Deacetylase ◽  
Targeted Inhibitors ◽  
In Cells

SIRT3 is a mitochondrial lysine deacetylase enzyme, regulating the activity of numerous mitochondrial proteins. Here, we have designed inhibitors of this enzyme, which exhibit selective inhibition of SIRT3 in cells by specific organelle localizing.

scholarly journals Mitochondria-Targeted Inhibitors of the Human SIRT3 Lysine Deacetylase

2020 ◽  
Author(s):  
Kathrin Sten Troelsen ◽  
Michael Bæk, ◽  
Alexander Lund Nielsen ◽  
Andreas Stahl Madasen ◽  
Nima Rajabi ◽  
...  
Keyword(s):  
Drug Target ◽  
Selective Inhibition ◽  
Pharmacological Intervention ◽  
Major Protein ◽  
Potential Drug Target ◽  
Potential Drug ◽  
Lysine Deacetylase ◽  
Wide Range ◽  
Acetylation State ◽  
In Cells

<p>Sirtuin 3 (SIRT3) is the major protein lysine deacetylase in the mitochondria. This hydrolase regulates a wide range of metabolically involved enzymes and has been considered as a potential drug target in certain cancers. Investigation of pharmacological intervention has been challenging due to a lack of potent and selective inhibitors of SIRT3. Here, we developed a strategy for selective inhibition of SIRT3 in cells, over its structurally similar isozymes that localize primarily to nucleus (SIRT1) and cytoplasm (SIRT2). This was achieved by directing the inhibitors straight to the mitochondria through incorporation of sequences inspired by previously described mitochondria-targeting peptides. Our inhibitors exhibited excellent mitochondrial localization in HeLa cells as indicated by fluorophore-conjugated versions and target engagement was demonstrated by a thermal shift assay of SIRT3 using western blotting. The acetylation state of documented SIRT3 target MnSOD was shown to be perturbed in cells with little effect on known targets of SIRT1 and SIRT2, showing that our lead compound exhibits selectivity for SIRT3 in cells. We expect that the developed inhibitor will now enable a more detailed investigation of SIRT3 as a potential drug target and help shed further light on the diverse biology regulated by this enzyme. </p>

scholarly journals Mitochondria-Targeted Inhibitors of the Human SIRT3 Lysine Deacetylase

2020 ◽  
Author(s):  
Kathrin Sten Troelsen ◽  
Michael Bæk, ◽  
Andreas Stahl Madasen ◽  
Nima Rajabi ◽  
Christian Adam Olsen
Keyword(s):  
Drug Target ◽  
Selective Inhibition ◽  
Pharmacological Intervention ◽  
Major Protein ◽  
Potential Drug Target ◽  
Potential Drug ◽  
Lysine Deacetylase ◽  
Wide Range ◽  
Acetylation State ◽  
In Cells

<p>Sirtuin 3 (SIRT3) is the major protein lysine deacetylase in the mitochondria. This hydrolase regulates a wide range of metabolically involved enzymes and has been considered as a potential drug target in certain cancers. Investigation of pharmacological intervention has been challenging due to a lack of potent and selective inhibitors of SIRT3. Here, we developed a strategy for selective inhibition of SIRT3 in cells, over its structurally similar isozymes that localize primarily to nucleus (SIRT1) and cytoplasm (SIRT2). This was achieved by directing the inhibitors straight to the mitochondria through incorporation of sequences inspired by previously described mitochondria-targeting peptides. Our inhibitors exhibited excellent mitochondrial localization in HeLa cells as indicated by fluorophore-conjugated versions and target engagement was demonstrated by a thermal shift assay of SIRT3 using western blotting. The acetylation state of documented SIRT3 target MnSOD was shown to be perturbed in cells with little effect on known targets of SIRT1 and SIRT2, showing that our lead compound exhibits selectivity for SIRT3 in cells. We expect that the developed inhibitor will now enable a more detailed investigation of SIRT3 as a potential drug target and help shed further light on the diverse biology regulated by this enzyme. </p>

scholarly journals Mitochondria-Targeted Inhibitors of the Human SIRT3 Lysine Deacetylase

2020 ◽  
Author(s):  
Kathrin Sten Troelsen ◽  
Michael Bæk, ◽  
Alexander Lund Nielsen ◽  
Andreas Stahl Madasen ◽  
Nima Rajabi ◽  
...  
Keyword(s):  
Drug Target ◽  
Selective Inhibition ◽  
Pharmacological Intervention ◽  
Major Protein ◽  
Potential Drug Target ◽  
Potential Drug ◽  
Lysine Deacetylase ◽  
Wide Range ◽  
Acetylation State ◽  
In Cells

<p>Sirtuin 3 (SIRT3) is the major protein lysine deacetylase in the mitochondria. This hydrolase regulates a wide range of metabolically involved enzymes and has been considered as a potential drug target in certain cancers. Investigation of pharmacological intervention has been challenging due to a lack of potent and selective inhibitors of SIRT3. Here, we developed a strategy for selective inhibition of SIRT3 in cells, over its structurally similar isozymes that localize primarily to nucleus (SIRT1) and cytoplasm (SIRT2). This was achieved by directing the inhibitors straight to the mitochondria through incorporation of sequences inspired by previously described mitochondria-targeting peptides. Our inhibitors exhibited excellent mitochondrial localization in HeLa cells as indicated by fluorophore-conjugated versions and target engagement was demonstrated by a thermal shift assay of SIRT3 using western blotting. The acetylation state of documented SIRT3 target MnSOD was shown to be perturbed in cells with little effect on known targets of SIRT1 and SIRT2, showing that our lead compound exhibits selectivity for SIRT3 in cells. We expect that the developed inhibitor will now enable a more detailed investigation of SIRT3 as a potential drug target and help shed further light on the diverse biology regulated by this enzyme. </p>

Specific and selective inhibition of translation of mRNA for type II procollagen in cells of transgenic mice that synthesize type I procollagen. A possible role for endogneous antisense RNA

1996 ◽  
Vol 15 (3) ◽  
pp. 203-210
Author(s):  
Leena Ala-Kokko ◽  
Constance Yuan ◽  
Dominique Le Guellec ◽  
Suzanne Franc ◽  
Andrzej Fertala ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Antisense Rna ◽  
Selective Inhibition ◽  
Type I ◽  
Type Ii ◽  
Type I Procollagen ◽  
In Cells

scholarly journals Expression of Mutant Dynamin Inhibits Toxicity and Transport of Endocytosed Ricin to the Golgi Apparatus

1998 ◽  
Vol 140 (3) ◽  
pp. 553-563 ◽  
Author(s):  
Alicia Llorente ◽  
Andrzej Rapak ◽  
Sandra L. Schmid ◽  
Bo van Deurs ◽  
Kirsten Sandvig
Keyword(s):  
Golgi Apparatus ◽  
Intracellular Transport ◽  
Trichostatin A ◽  
Selective Inhibition ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Tyrosine Sulfation ◽  
Cell Biol ◽  
The Difference ◽  
In Cells

Endocytosis and intracellular transport of ricin were studied in stable transfected HeLa cells where overexpression of wild-type (WT) or mutant dynamin is regulated by tetracycline. Overexpression of the temperature-sensitive mutant dynG273D at the nonpermissive temperature or the dynK44A mutant inhibits clathrin-dependent endocytosis (Damke, H., T. Baba, A.M. van der Blieck, and S.L. Schmid. 1995. J. Cell Biol. 131: 69–80; Damke, H., T. Baba, D.E. Warnock, and S.L. Schmid. 1994. J. Cell Biol. 127:915–934). Under these conditions, ricin was endocytosed at a normal level. Surprisingly, overexpression of both mutants made the cells less sensitive to ricin. Butyric acid and trichostatin A treatment enhanced dynamin overexpression and increased the difference in toxin sensitivity between cells with normal and mutant dynamin. Intoxication with ricin seems to require toxin transport to the Golgi apparatus (Sandirg, K., and B. van Deurs. 1996. Physiol. Rev. 76:949–966), and this process was monitored by measuring the incorporation of radioactive sulfate into a modified ricin molecule containing a tyrosine sulfation site. The sulfation of ricin was much greater in cells expressing dynWT than in cells expressing dynK44A. Ultrastructural analysis using a ricin-HRP conjugate confirmed that transport to the Golgi apparatus was severely inhibited in cells expressing dynK44A. In contrast, ricin transport to lysosomes as measured by degradation of 125I-ricin was essentially unchanged in cells expressing dynK44A. These data demonstrate that although ricin is internalized by clathrin-independent endocytosis in cells expressing mutant dynamin, there is a strong and apparently selective inhibition of ricin transport to the Golgi apparatus. Also, in cells with mutant dynamin, there is a redistribution of the mannose-6-phosphate receptor.

scholarly journals GSK3 Kinase Inhibitor, CHIR, Suppress Transcription of Tissue Specific POU2F1 Isoform in Burkitt Namalwa Lymphoma Cells

2021 ◽  
Vol 496 (1) ◽  
pp. 32-35
Author(s):  
E. V. Pankratova ◽  
T. N. Portseva ◽  
A. A. Makarova ◽  
Yu. V. Ilyin ◽  
A. G. Stepchenko ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Tumor Cells ◽  
Kinase Inhibitor ◽  
Malignant Tumors ◽  
B Cell Lymphoma ◽  
Selective Inhibition ◽  
Alternative Promoters ◽  
Tissue Specific ◽  
Human Malignant Tumors ◽  
In Cells

Abstract POU2F1 (Oct-1) is a transcription factor, the overexpression of which is found in many human malignant tumors; a significant increase in its level in cells determines the malignant potential of the tumor. POU2F1 is represented in cells by several isoforms that are transcribed from alternative promoters. In Burkitt’s B-cell lymphoma Namalwa, the concentration of tissue-specific isoform Oct-1L is several times higher than in normal B cells. We tested the potential to inhibit the transcription of individual Oct-1 isoforms using the GSK3 kinase inhibitor CHIR, an aminopyrimidine derivative. We have shown that CHIR specifically affects the expression of the tissue-specific isoform Oct-1L, significantly reducing the level of mRNA and Oct-1L protein. However, CHIR does not change the amount of mRNA and protein of the ubiquitous isoform Oct-1A in Namalwa tumor cells. The results obtained show that it is possible to develop a system for selective inhibition of Oct-1 transcription factor isoforms in human cells to suppress drug resistance of tumor cells with a high POU2F1 content.

scholarly journals Loss of mitochondrial hsp60 function: nonequivalent effects on matrix-targeted and intermembrane-targeted proteins.

1993 ◽  
Vol 13 (5) ◽  
pp. 3050-3057 ◽  
Author(s):  
E M Hallberg ◽  
Y Shu ◽  
R L Hallberg
Keyword(s):  
Mitochondrial Proteins ◽  
Temperature Sensitive ◽  
Intermembrane Space ◽  
Mitochondrial Matrix ◽  
Nonpermissive Temperature ◽  
Yeast Strains ◽  
Cytochrome C1 ◽  
Normal Processing ◽  
Correct Processing ◽  
In Cells

We have created yeast strains in which the mitochondrial chaperonin, hsp60, can be either physically depleted or functionally inactivated. Cells completely depleted of hsp60 stop growing but retain for awhile the capacity to reaccumulate hsp60. While this newly made hsp60 is targeted to and processed correctly within the mitochondrion, assembly of a functional hsp60 complex does not occur. Rather, the hsp60 monomers are localized in different-size soluble complexes containing another mitochondrial chaperone, the mitochondrial form of hsp70. A number of other mitochondrial matrix-targeted proteins synthesized in the absence of functional hsp60 are imported into mitochondria but often show some buildup of precursor forms and, unlike hsp60, accumulate as insoluble aggregates. By contrast, several mitochondrial proteins normally targeted to the intermembrane space show normal processing in the complete absence of a functional hsp60 complex. Similar and complementary results were obtained when we examined the metabolism of matrix- and intermembrane space-localized proteins in cells expressing three different temperature-sensitive alleles of HSP60. In all cases, matrix-targeted proteins synthesized at nonpermissive (i.e., hsp60-inactivating) temperatures were correctly targeted to and processed within mitochondria but accumulated predominantly or totally as insoluble aggregates. The metabolism of two intermembrane space proteins, cytochrome b2 and cytochrome c1, was unaffected at the nonpermissive temperature, as judged by the correct processing and complete solubility of newly synthesized forms of both proteins. These findings are discussed with regard to current models of intermembrane targeting.

Selective Inhibition of a Nuclear RNA Polymerase in Cells infected with Frog Virus

Nature ◽  
1970 ◽  
Vol 227 (5255) ◽  
pp. 294-295 ◽  
Author(s):  
F. COSTANZO ◽  
M. LA PLACA ◽  
ANNA MANNINI PALENZONA ◽  
F. NOVELLO ◽  
F. STIRPE
Keyword(s):  
Rna Polymerase ◽  
Selective Inhibition ◽  
Frog Virus ◽  
Nuclear Rna ◽  
Nuclear Rna Polymerase ◽  
In Cells

scholarly journals Induction of mitochondrial proteins in MA-10 Leydig tumour cells with human choriogonadotropin

1988 ◽  
Vol 249 (1) ◽  
pp. 95-103 ◽  
Author(s):  
D M Stocco ◽  
M W Kilgore
Keyword(s):  
Cyclic Amp ◽  
Polyacrylamide Gel Electrophoresis ◽  
Tumour Cells ◽  
Mitochondrial Proteins ◽  
Rapid Induction ◽  
Human Choriogonadotropin ◽  
Chain Cleavage ◽  
Specific Proteins ◽  
Stimulation Of ◽  
In Cells

The side-chain cleavage of cholesterol is the rate-limiting enzymic step in steroidogenesis and occurs in the mitochondria of steroid-producing tissues. In these studies, the effect of acute stimulation of MA-10 Leydig-tumour cells on the synthesis of mitochondrial proteins was investigated. Cells were incubated in the presence of stimulating levels of human choriogonadotropin (hCG) and [35S]methionine for 2 h periods. Mitochondria were isolated and their proteins analysed by two-dimensional polyacrylamide-gel electrophoresis and fluorography. At least three mitochondrial-specific proteins were found in cells exposed to gonadotropin, and these proteins were also found in cells treated with dibutyryl cyclic AMP (db cyclic AMP). The appearance of these proteins was prevented by the addition of cycloheximide to the cells, as was the production of progesterone, the major steroid produced in MA-10 cells. In addition, mitochondria isolated from cells stimulated with db cyclic AMP produced progesterone at a rate 3-fold greater than mitochondria isolated from control cells during 3 h of incubation. Lastly, mixing experiments demonstrated that sonicated mitochondria isolated from db cyclic AMP-treated cells stimulated progesterone production in control mitochondria 6-fold. These studies show that hCG and db cyclic AMP stimulation of MA-10 cells results in the rapid induction of cycloheximide-sensitive proteins located in the mitochondria which may be instrumental in the acute regulation of steroidogenesis.

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