Division of Molecular Biology
Research Summary
Our main research aimed at analyses on the relationship and regulation mechanisms of protein degradation systems controlled by nutrient and environmental conditions.
Research Project
Aging and Cell Death
The proliferating cells are finally fated to fall in apoptosis, senescence, and tumorigenesis. In our laboratory, we identified some proteins interacting caspase-3, an executioner of apoptosis. We investigated on the roles of these proteins in apoptosis and senescence.
Senescence marker protein-30 (SMP30) interacted with procaspase-3 but not active-caspase-3. Amount of SMP30 decreased with aging, which is identical with -gluconolactonase participated in vitamin C synthesis. SMP30 affected the cellular sensitivity to apoptosis inducers; Up-regulation of SMP30 increased cellular sensitivity to apoptosis inducers, whereas downregulation decreased them. Amount of SMP30 decreased in most of cancerous tissues. Thus, our laboratory investigated the inducer of SMP30 in cancerous tissues which is a promised agent for novel anti-cancer therapy.
Quinolinate phosphoribosyl transferase (QPRT) interacted with active-caspase-3 but not procaspase-3. QPRT-depleted cells possessed large aggregates of active-caspase-3 resulting in spontaneous cell death. QPRT participated in de nove NAD+ synthesis from tryptophan in liver and kidney. Amount of QPRT decreased in aging cells, whereas those increased in most of tumor tissues.
Tryptophan metabolism closely related intracellular energy production through de novo NAD synthesis. Some intermediates in this pathway bind the receptor for arylhydrocarbon, which was identified as environmental hormones. Upon the activation of this receptor system, immune cells suppress proliferation, neural cells suppress apoptosis, and tumor cells stimulate proliferation. N-(3,4-Dimethoxycinnamoyl) anthronic acid (3,4-DAA), a synthetic analogue of 3-hydroxyanthronia acid or 3-hydorxykynurenine inhibits the arylhydrocarbone receptor pathway.
Caspase-7 seems to be one of the executioner caspases because of its amino acid sequence similarity to that of caspase-3. Substrate specificity of caspase-7 is closely related to that of caspase-3, both caspases cleaves DEVD-sequence. Now, we develop the caspase-7-specific FRET vector detecting intracellular caspase-7 activity in the living cells as well as caspase-7-specific inhibitor.
Selected Papers
1. H-L Zhang, Y Luo, X-Y Zhou, J-G Rong, Y Ni, M L, K Ishidoh, Z-H Liu, T A. Kent, Y-M Zhu, W Li, and S-G Qiao: Inhibition of autophagy blocks cathepsins-tBid-mitochondrial apoptotic signaling pathway via stabilization of lysosomal membrane in ischemic astrocytes" Cell Death Disease 8(2):e2618 2017
2. Kamemura N, Murakami S, Komatsu H, Sawanoi M, Miyamoto K, Ishidoh K, Kishimoto K, Tsuji A, Yuasa K Type II cGMP-dependent protein kinase negatively regulates fibroblast growth factor signaling by phosphorylating Raf-1 at serine 43 in rat chondrosarcoma cells. Biochem Biophys Res Commun. 483:82-87 2017
3. Xu M, Yang L, Rong JG, Ni Y, Gu WW, Luo Y, Ishidoh K, Katunuma N, Li ZS, Zhang HL. Inhibition of cysteine cathepsin B and L activation in astrocytes contributes to neuroprotection against cerebral ischemia via blocking the tBid-mitochondrial apoptotic signaling pathway.
Glia. 62:855-80.2014
4. Nagahama M, Takahashi C, Aoyanagi K, Tashiro R, Kobayashi K, Sakaguchi Y, Ishidoh K, Sakurai J. Intracellular trafficking of Clostridium botulinum C2 toxin. Toxicon. 82:76-82. 2014
5. Gu WW, Ao GZ, Zhu YM, Sun SC, Zhou Q, Fan JH, Nobuhiko K, Ishidoh K, Zhang HL, Gao XM. Autophagy and cathepsin L are involved in the antinociceptive effect of DMBC in a mouse acetic acid-writhing model.Acta Pharmacol Sin. 34, 1007-1012. 2013
6. Inubushi T, Kamemura N, Oda M, Sakurai J, Nakaya Y, Harada N, Suenaga M, Matsunaga Y, Ishidoh K, Katunuma N. L-tryptophan suppresses blood-glucose elevation and stimulates glucose-energy expenditure in rats. J Nutritional Sci and Vitaminol. 58, 415-422. 2012
7. Nagahama M, Umezaki M, Tashiro R, Oda M, Kobayashi K, Shibutani M, Takagishi T, Ishidoh K, Fukuda M, Sakurai J. Intracellular Trafficking of Clostridium perfringens Iota-Toxin b. Infec Immunity 80, 3410-3416.2012
8. Imagawa T, Tsurumura T, Sugimoto Y, Aki K, Ishidoh K, Kuramitsu S, Tsuge H. :Structural basis of free reduced flavin generation by flavin reductase from Thermus thermophilus HB8. J Biol Chem 286, 44078-44085.2011
9. Nagahama M, Umezaki M, Oda M Kobayashi K, Tone S, Ishidoh K, Sakurai J: Clostridium perfringens iota-b induces rapid cell necrosis. Infec. Immun. 79, 4353-4360. 2011
10. Matsugaki T, Shiba N, Kohno S, Nikawa T, Hirasaka K, Okumura Y, Ishidoh K, Soejima T, Yoshimitsu K, Nagata K.:Hybrid exercise" prevents muscle atrophy in association with a distinct gene expression pattern.Kurume Med J. 57(4):101-108. 2011
11. Sugita S, Yamada Y, Horie S, Nakamura O, Ishidoh K, Yamamoto Y, Yamagami S, Mochizuki M.:Induction of T regulatory cells by cytotoxic T-lymphocyte antigen-2α on corneal endothelial cells.Invest Ophthalmol Vis Sci. 52(5):2598-2605.2011
12. Kido, H and Ishidoh K:Nobuhiko Katunuma: An Outstanding Scientist in the field of Proteolysis and Warm-Hearted 'Kendo Fighter' Biochemist. J. Biochem. 148:527-531.2010
13. Ishidoh K, Kamemura N, Imagawa T, Oda M, Sakurai J, Katunuma N: Quinolinate Phosphoribosyl Transferase. a key enzyme in de novo NAD+ synthesis, suppress spontenous cell death by inhibiting overproduction of active-caspase-3. Biochim Biophys Acta 1803; 527-533. 2010
14. Sugita S, Horie S, Nakamura O, Maruyama K, Takase H, Usui Y, Takeuchi M, Ishidoh K, Koike M, Uchiyama Y, Peters C, Yamamoto Y, Mochizuki M: Acquisition of T regulatory function in Cathepsin L-inhibited T cells by eye-derived CTLA-2a during inflammatory conditions. J Immunol 183 (10), 5013-5022. 2009
15. Nakao R, Hirasaka K, Goto J, Ishidoh K, Yamada C, Ohno A, Okumura Y, Nonaka I, Yasutomo K, Baldwin KM, Kominami E, Higashibata A, Nagano K, Tanaka K, Yasui N, Mills EM, Takeda S, Nikawa T.: Ubiquitin ligase Cbl-b is a negative regulator for IGF-1 signaling during muscle atrophy caused by unloading. Mol Cell Biol. 29(17):4798-811.2009
16. Ogawa N, Takahashi M, Ishidoh K, Katunuma N.: Inhibition of collagenolytic cathepsins by beta-lactoglobulin in milk and its suppressive effect on bone resorption. J Nutr Sci Vitaminol (Tokyo). 55(3):264-70. 2009
17. Sugita S, Horie S, Nakamura O, Futagami Y, Takase H, Keino H, Aburatani H, Katunuma N, Ishidoh K, Yamamoto Y, Mochizuki M. Retinal pigment epithelium-derived CTLA-2alpha induces TGFbeta-producing T regulatory cells.
J Immunol. 181(11):7525-7536. 2008
18. Hirasaka K, Tokuoka N, Nakao R, Yamada C, Oarada M, Imagawa T, Ishidoh K, Okumura Y, Kishi K, Nikawa T: Cathepsin C propeptide interacts with intestinal alkaline phosphatase and heat shock cognate protein 70 in human Caco-2 cells. J Physiol Sci 58, 105-111. 2008
19. Furochi H, Tamura S, Mameoka M, Yamada C, Ogawa T, Hirasaka K, Okumura Y, Oguri S, Ishidoh K, Kishi K, Higashiyama S, Nikawa T. : Osteoactivin fragments produced by ectodomain shedding induce MMP-3 expression via ERK pathway in mouse NIH3T3 fibroblasts. FEBS Lett. 581, 5743-5750. 2007
20. Hirasaka K., Kohno S., Goto J., Furochi H., Mawatari K., Harada N., Hosaka T., Nakaya Y., Ishidoh K., Obata T., Ebina Y., Gu H., Takeda S., Kishi K., Nikawa T.: Deficiency of Cbl-b gene enhances infiltration and activation of macrophages in adipose tissue and causes peripheral insulin resistance in mice. Diabetes 56, 2511-2522. 2007
21. Ogawa T, Furochi H, Mameoka M, Hirasaka K, Onishi Y, Suzue N, Oarada M, Akamatsu M, Akima, H, Fukunaga T, Kishi K, Yasui N, Ishidoh K, Fukuoka H, Nikawa T.: Ubiquitin ligase gene expression in healthy volenteers with 20-day bed rest. Muscle Nerve 34,463-469.2006.
22. Suzue N, Nikawa T, Onishi Y, Yamada, C, Hirasaka K, Ogawa T, Furochi H, Kosaka H, Ishidoh K, Gu H, Takeda S, Ishimaru N, Hayashi Y, Yamamoto H, Kishi K Yasui N: Ubiquitinligase Cbl-b downregulates bone formation through suppression of IGF-I signaling in osteoblasts during denervation. J. Bone Mineral Res. 21, 722-734. 2006
23. Hashimoto M, Nakajima-Shimada J, Ishidoh K, Aoki T.: Gene expression profiles in response to Fas stimulation in Trypanosoma cruzi-infected host cells. Int. J. Parasitol. 25, 1587-1594, 2005
24. Onishi Y, Hirasaka K, Ishihara I, Oarada M, Goto J, Ogawa T, Suzue N, Nakano S, Furochi H, Ishidoh K, Kishi K, Nikawa T.: Identification of mono-ubiquitinated LDH-A in skeletal muscle cells exposed to oxidative stress. Biochem. Biophys. Res. Commun. 336, 799-806. 2005.
Research Summary
Our main research aimed at analyses on the relationship and regulation mechanisms of protein degradation systems controlled by nutrient and environmental conditions.
Research Project
Aging and Cell Death
The proliferating cells are finally fated to fall in apoptosis, senescence, and tumorigenesis. In our laboratory, we identified some proteins interacting caspase-3, an executioner of apoptosis. We investigated on the roles of these proteins in apoptosis and senescence.
Senescence marker protein-30 (SMP30) interacted with procaspase-3 but not active-caspase-3. Amount of SMP30 decreased with aging, which is identical with -gluconolactonase participated in vitamin C synthesis. SMP30 affected the cellular sensitivity to apoptosis inducers; Up-regulation of SMP30 increased cellular sensitivity to apoptosis inducers, whereas downregulation decreased them. Amount of SMP30 decreased in most of cancerous tissues. Thus, our laboratory investigated the inducer of SMP30 in cancerous tissues which is a promised agent for novel anti-cancer therapy.
Quinolinate phosphoribosyl transferase (QPRT) interacted with active-caspase-3 but not procaspase-3. QPRT-depleted cells possessed large aggregates of active-caspase-3 resulting in spontaneous cell death. QPRT participated in de nove NAD+ synthesis from tryptophan in liver and kidney. Amount of QPRT decreased in aging cells, whereas those increased in most of tumor tissues.
Tryptophan metabolism closely related intracellular energy production through de novo NAD synthesis. Some intermediates in this pathway bind the receptor for arylhydrocarbon, which was identified as environmental hormones. Upon the activation of this receptor system, immune cells suppress proliferation, neural cells suppress apoptosis, and tumor cells stimulate proliferation. N-(3,4-Dimethoxycinnamoyl) anthronic acid (3,4-DAA), a synthetic analogue of 3-hydroxyanthronia acid or 3-hydorxykynurenine inhibits the arylhydrocarbone receptor pathway.
Caspase-7 seems to be one of the executioner caspases because of its amino acid sequence similarity to that of caspase-3. Substrate specificity of caspase-7 is closely related to that of caspase-3, both caspases cleaves DEVD-sequence. Now, we develop the caspase-7-specific FRET vector detecting intracellular caspase-7 activity in the living cells as well as caspase-7-specific inhibitor.
Selected Papers
1. H-L Zhang, Y Luo, X-Y Zhou, J-G Rong, Y Ni, M L, K Ishidoh, Z-H Liu, T A. Kent, Y-M Zhu, W Li, and S-G Qiao: Inhibition of autophagy blocks cathepsins-tBid-mitochondrial apoptotic signaling pathway via stabilization of lysosomal membrane in ischemic astrocytes" Cell Death Disease 8(2):e2618 2017
2. Kamemura N, Murakami S, Komatsu H, Sawanoi M, Miyamoto K, Ishidoh K, Kishimoto K, Tsuji A, Yuasa K Type II cGMP-dependent protein kinase negatively regulates fibroblast growth factor signaling by phosphorylating Raf-1 at serine 43 in rat chondrosarcoma cells. Biochem Biophys Res Commun. 483:82-87 2017
3. Xu M, Yang L, Rong JG, Ni Y, Gu WW, Luo Y, Ishidoh K, Katunuma N, Li ZS, Zhang HL. Inhibition of cysteine cathepsin B and L activation in astrocytes contributes to neuroprotection against cerebral ischemia via blocking the tBid-mitochondrial apoptotic signaling pathway.
Glia. 62:855-80.2014
4. Nagahama M, Takahashi C, Aoyanagi K, Tashiro R, Kobayashi K, Sakaguchi Y, Ishidoh K, Sakurai J. Intracellular trafficking of Clostridium botulinum C2 toxin. Toxicon. 82:76-82. 2014
5. Gu WW, Ao GZ, Zhu YM, Sun SC, Zhou Q, Fan JH, Nobuhiko K, Ishidoh K, Zhang HL, Gao XM. Autophagy and cathepsin L are involved in the antinociceptive effect of DMBC in a mouse acetic acid-writhing model.Acta Pharmacol Sin. 34, 1007-1012. 2013
6. Inubushi T, Kamemura N, Oda M, Sakurai J, Nakaya Y, Harada N, Suenaga M, Matsunaga Y, Ishidoh K, Katunuma N. L-tryptophan suppresses blood-glucose elevation and stimulates glucose-energy expenditure in rats. J Nutritional Sci and Vitaminol. 58, 415-422. 2012
7. Nagahama M, Umezaki M, Tashiro R, Oda M, Kobayashi K, Shibutani M, Takagishi T, Ishidoh K, Fukuda M, Sakurai J. Intracellular Trafficking of Clostridium perfringens Iota-Toxin b. Infec Immunity 80, 3410-3416.2012
8. Imagawa T, Tsurumura T, Sugimoto Y, Aki K, Ishidoh K, Kuramitsu S, Tsuge H. :Structural basis of free reduced flavin generation by flavin reductase from Thermus thermophilus HB8. J Biol Chem 286, 44078-44085.2011
9. Nagahama M, Umezaki M, Oda M Kobayashi K, Tone S, Ishidoh K, Sakurai J: Clostridium perfringens iota-b induces rapid cell necrosis. Infec. Immun. 79, 4353-4360. 2011
10. Matsugaki T, Shiba N, Kohno S, Nikawa T, Hirasaka K, Okumura Y, Ishidoh K, Soejima T, Yoshimitsu K, Nagata K.:Hybrid exercise" prevents muscle atrophy in association with a distinct gene expression pattern.Kurume Med J. 57(4):101-108. 2011
11. Sugita S, Yamada Y, Horie S, Nakamura O, Ishidoh K, Yamamoto Y, Yamagami S, Mochizuki M.:Induction of T regulatory cells by cytotoxic T-lymphocyte antigen-2α on corneal endothelial cells.Invest Ophthalmol Vis Sci. 52(5):2598-2605.2011
12. Kido, H and Ishidoh K:Nobuhiko Katunuma: An Outstanding Scientist in the field of Proteolysis and Warm-Hearted 'Kendo Fighter' Biochemist. J. Biochem. 148:527-531.2010
13. Ishidoh K, Kamemura N, Imagawa T, Oda M, Sakurai J, Katunuma N: Quinolinate Phosphoribosyl Transferase. a key enzyme in de novo NAD+ synthesis, suppress spontenous cell death by inhibiting overproduction of active-caspase-3. Biochim Biophys Acta 1803; 527-533. 2010
14. Sugita S, Horie S, Nakamura O, Maruyama K, Takase H, Usui Y, Takeuchi M, Ishidoh K, Koike M, Uchiyama Y, Peters C, Yamamoto Y, Mochizuki M: Acquisition of T regulatory function in Cathepsin L-inhibited T cells by eye-derived CTLA-2a during inflammatory conditions. J Immunol 183 (10), 5013-5022. 2009
15. Nakao R, Hirasaka K, Goto J, Ishidoh K, Yamada C, Ohno A, Okumura Y, Nonaka I, Yasutomo K, Baldwin KM, Kominami E, Higashibata A, Nagano K, Tanaka K, Yasui N, Mills EM, Takeda S, Nikawa T.: Ubiquitin ligase Cbl-b is a negative regulator for IGF-1 signaling during muscle atrophy caused by unloading. Mol Cell Biol. 29(17):4798-811.2009
16. Ogawa N, Takahashi M, Ishidoh K, Katunuma N.: Inhibition of collagenolytic cathepsins by beta-lactoglobulin in milk and its suppressive effect on bone resorption. J Nutr Sci Vitaminol (Tokyo). 55(3):264-70. 2009
17. Sugita S, Horie S, Nakamura O, Futagami Y, Takase H, Keino H, Aburatani H, Katunuma N, Ishidoh K, Yamamoto Y, Mochizuki M. Retinal pigment epithelium-derived CTLA-2alpha induces TGFbeta-producing T regulatory cells.
J Immunol. 181(11):7525-7536. 2008
18. Hirasaka K, Tokuoka N, Nakao R, Yamada C, Oarada M, Imagawa T, Ishidoh K, Okumura Y, Kishi K, Nikawa T: Cathepsin C propeptide interacts with intestinal alkaline phosphatase and heat shock cognate protein 70 in human Caco-2 cells. J Physiol Sci 58, 105-111. 2008
19. Furochi H, Tamura S, Mameoka M, Yamada C, Ogawa T, Hirasaka K, Okumura Y, Oguri S, Ishidoh K, Kishi K, Higashiyama S, Nikawa T. : Osteoactivin fragments produced by ectodomain shedding induce MMP-3 expression via ERK pathway in mouse NIH3T3 fibroblasts. FEBS Lett. 581, 5743-5750. 2007
20. Hirasaka K., Kohno S., Goto J., Furochi H., Mawatari K., Harada N., Hosaka T., Nakaya Y., Ishidoh K., Obata T., Ebina Y., Gu H., Takeda S., Kishi K., Nikawa T.: Deficiency of Cbl-b gene enhances infiltration and activation of macrophages in adipose tissue and causes peripheral insulin resistance in mice. Diabetes 56, 2511-2522. 2007
21. Ogawa T, Furochi H, Mameoka M, Hirasaka K, Onishi Y, Suzue N, Oarada M, Akamatsu M, Akima, H, Fukunaga T, Kishi K, Yasui N, Ishidoh K, Fukuoka H, Nikawa T.: Ubiquitin ligase gene expression in healthy volenteers with 20-day bed rest. Muscle Nerve 34,463-469.2006.
22. Suzue N, Nikawa T, Onishi Y, Yamada, C, Hirasaka K, Ogawa T, Furochi H, Kosaka H, Ishidoh K, Gu H, Takeda S, Ishimaru N, Hayashi Y, Yamamoto H, Kishi K Yasui N: Ubiquitinligase Cbl-b downregulates bone formation through suppression of IGF-I signaling in osteoblasts during denervation. J. Bone Mineral Res. 21, 722-734. 2006
23. Hashimoto M, Nakajima-Shimada J, Ishidoh K, Aoki T.: Gene expression profiles in response to Fas stimulation in Trypanosoma cruzi-infected host cells. Int. J. Parasitol. 25, 1587-1594, 2005
24. Onishi Y, Hirasaka K, Ishihara I, Oarada M, Goto J, Ogawa T, Suzue N, Nakano S, Furochi H, Ishidoh K, Kishi K, Nikawa T.: Identification of mono-ubiquitinated LDH-A in skeletal muscle cells exposed to oxidative stress. Biochem. Biophys. Res. Commun. 336, 799-806. 2005.
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