Division of Enzyme Chemistry

Main Research AreasResearch ProjectsMemberPublications


Main Research Areas

Studies on the pathogenesis of diseases and therapeutics by the analyses of enzymology, proteomics and metabolome.


Studies on the pathogenesis of infectious diseases and prevention by mucosal immunity.

  • Influenza virus hemagglutinin (HA) processing proteases which proteolytically activate seasonal influenza virus and highly pathogenic avian influenza virus entry.
  • Pathogenesis of influenza-associated encephalopathy and multiple organ failure evoked by influenza virus infection.
  • Development of mucosal influenza vaccine.


Studies on ATPase and nucleoside diphosphate kinase activities of molecular chaperones, such as Hsp70 and 14-3-3 proteins. Role of stress-induced 14-3-3 proteins in apoptosis of cancer cells.


Development of diamond-like carbon coated protein tips for detection of antibodies with high sensitivity and application of these protein tips for clinical diagnosis of food allergy.


Research Projects

1.Studies on the pathogenesis of influenza virus entry.

Studies on the pathogenesis of influenza virus entry.
< Fig.1 Replication cycle of influenza virus >

Proteolytic processing of Influenza virus HA is indispensable and prerequisite for virus entry into cells. We first identified mammalian HA processing enzyme, tryptase Clara, from the lungs and have also identified series of trypsin-type processing proteases in the airway for seasonal influenza viruses and highly pathogenic avian influenza viruses. These proteases might be drug targets which inhibit virus entry.


2.Studies on the pathogenesis of influenza-associated encephalopathy and multiple organ failure.

Studies on the pathogenesis of influenza-associated encephalopathy and multiple organ failure.
< Fig. 2 IAE and multiple-organ failure
evoked by influenza virus infection >

Influenza-associated encephalopathy (IAE) is a pediatric complication of influenza virus infection characterized by the sudden onset of febrile convelsions and multiple-organ failure during high fever. We have identified genetic factors which may play an important role in the etiology of IAE. In a surveillance study initiated in the year 2000 in Japan, we identified disease-causing mutations and compound variations of mitochondrial enzymes in the died or disabled patients: thermal instability of compound variants of carnitine palmitoyltransferase II (CPT 2) and mutations of several enzymes for ATP generation.


Studies on the pathogenesis of influenza-associated encephalopathy and multiple organ failure.02
< Fig.3 Metabolic energy generation in mitochondria >

Since the intracellular depletion of ATP in the IAE patients causes disassembly of extracellular tight junctions of endothelial cells, restoration of ATP levels might be an effective treatment of IAE. We are studying on the metabolic treatments for restoration of ATP.




3.Pulmonary surfactant is a natural adjuvant for mucosal immunity.

Pulmonary surfactant is a natural adjuvant for mucosal immunity.
< Fig.4 Modified pulmonary surfactant
is a potent mucosal adjuvant >

We found that pulmonary surfactant in the lungs is a natural and potent mucosal adjuvant which effectively deliver antigen to dendritic cells. Based on this evidence, we are developing synthetic adjuvant imicking human pulmonary surfactant for intranasal administration of mucosal influenza vaccine. Currently available subcutaneous or intramuscular vaccines can only induce IgG response, but do not induce the production of sufficient mucosal IgA to prevent initial steps of infection. By using the synthetic adjuvant, we are developing safe and effective mucosal adjuvant, particularly for children.


4.Studies on molecular chaperone.

Studies on molecular chaperone.
< Fig.5 Regulation of expression of stress
proteins by NSAIDs and their applications
to cancer therapy >

We have identified nucleoside diphosphate kinase activity instead of ATPase activity in molecular chaperones, such as Hsp70 and 14-3-3 proteins. The role of nucleoside diphosphate kinase activity in the functions of these proteins are under investigation. These stress proteins play an essential role in the retention of cell homeostasis. On the other hand, in various cancer cells which express higher stress proteins levels, this causes the acceleration of cancer cell growth and invasion. We recently found that nonsteroidal anti-inflammatory drugs (NSAIDs) significantly suppressed the expression of stress proteins in cancer cells. NSAIDs which inhibit stress protein expression exert anti-tumor activity via stress protein-dependent activation of mitochondrial apoptosis pathway in cancer cells and increase the sensitivity of anticancer drugs.


5.Diamond-like carbon protein chips and development of them for diagnosis of food allergy.

Diamond-like carbon protein chips and development of them for diagnosis of food allergy.
< Fig.6 Allergy diagnostic protein tips >

Diamond-like carbon protein tips immobilize extremely high doses of proteins, which enable high-sensitive detection. By using this new product, we have developed high-sensitive IgE detection system for food allergy. Small doses of blood (10 nL), nasal fluids (1 μL) and saliva (1 μL) are useful samples for monitoring IgE levels in allergic patients.




Professor Hiroshi Kido, Ph.D., M.D.Professor
Hiroshi Kido, Ph.D., M.D.
TEL : 088-633-7423
FAX : 088-633-7425
E-mail : kido@ier.tokushima-u.ac.jp
Assistant Professor Dai Mizuno, Ph.D.Assistant Professor
Dai Mizuno, Ph.D.
TEL : 088-633-7424
FAX : 088-633-9436
E-mail : dmizuno@ier.tokushima-u.ac.jp
日吉峰麗Assistant Professor
Mineyoshi Hiyoshi, Ph.D.
TEL : 088-633-9235
E-mail : hiyoshi@ier.tokushima-u.ac.jp
亀村典生Assistant Professor
Norio Kamemura, Ph.D.
TEL : 088-633-9436
E-mail : kamemura@ier.tokushima-u.ac.jp





  1. Y. Cissé, S. Wang, I. Inoue and H. Kido: Rat model of influenza-associated encephalopathy (IAE): studies of electroencephalogram (EEG) in vivo, Neuroscience, 165, 1127-1137 (2010)
  2. Yiu-Wing Kam, Yuushi Okumura, Hiroshi Kido, Lisa F. P. Ng, Roberto Bruzzone, Ralf Altmeyer: Cleavage of the SARS coronavirus spike glycoprotein by airway proteases enhances virus entry into human bronchial epithelial cells in vitro, PLoS One, Nov 17, 4 (11), e7870 (2009)
  3. Takako Sawabuchi, Satoshi Suzuki, Kazuhiro Isase, Chika Ito, Dai Mizuno, Hajime Todari, Isamu Watanabe, Sadiqur R. Talukder, Junji Chida and Hiroshi Kido : Boost of mucosal secretory immunoglobulin A response by clarithromycin in paediatric influenza, Respirology, 14(8), 1173-1179 (2009)
  4. Maki Nishino, Dai Mizuno, Takashi Kimoto, Wakako Shinahara, Akiho Fukuta, Tsunetomo Takei, Kaori Sumida, Seiichiro Kitamura, Hiroshi Shiota , Hiroshi Kido: Influenza vaccine with Surfacten, a modified pulmonary surfactant, induces systemic and mucosal immune responses without side effects in minipigs, Vaccine, 27, 5620-5627 (2009)
  5. Hiroshi Kido, Yuushi Okumura, Etsuhisa Takahashi, Hai-Yan Pan, Siye Wang, Junji Chida, Trong Quang Le and Mihiro Yano: Host envelope glycoprotein processing proteases are indispensable for entry into human cells by seasonal and highly pathogenic avian influenza viruses, J. Mol. Genet. Med., 3(1), 167-175 (2009)
  6. Chisa Fujimoto, Hiroshi Kido, Takako Sawabuchi, Dai Mizuno, Masaki Hayama, Hiroaki Yanagawa and Noriaki Takeda: Evaluation of nasal IgA secretion in normal subjects by nasal spray and aspiration, Auris Nasus Larynx, in press (2009)
  7. Mineyoshi Hiyoshi, Hiroaki Konishi, Hirokazu Uemura, Hideki Matsuzaki, Hideo Tsukamoto, Ryusuke Sugimoto, Hideo Takeda, Satoru Takeshita, Asushi Kitayama, Hidenobu Takami, Fusakazu Sawachika, Hiroshi Kido and Kokichi Arisawa: d-Dopachrome tautomerase is a candidate for key proteins to protect the rat liver damaged by carbon tetrachloride. Toxicology, 255, 6-14 (2009)
  8. Daisuke Hashimoto, Masaki Ohmuraya, Masahiko Hirota, Akitsugu Yamamoto, Koichi Suyama, Satoshi Ida, Yuushi Okumura, Etsuhisa Takahashi, Hiroshi Kido, Kimi Araki, Hideo Baba, Noboru Mizushima, Ken-ichi Yamamura:Involvement of autophagy in trypsinogen activation within the pancreatic acinar cells, The Journal of Cell Biology, 18(7): 1065-1072 (2008)
  9. Shinichi Nakamuta, Hiroshi Endo, Youichiro Higashi, Aoi Kousaka, Hiroshi Yamada, Mihiro Yano and Hiroshi Kido:
    HIV-1 gp120-mediated disruption of tight junction proteins by induction of proteasome-mediated degradation of zonula occludens-1 and -2 in human brain microvascular endothelial cells, Journal of NeuroVirology, 14, 186-195 (2008)
  10. Dengbing Yao, Hiroshi Mizuguchi, Miyoko Yamaguchi, Hiroshi Yamada, Junji Chida, Koji Shikata and Hiroshi Kido: Thermal Instability of Compound Variants of Carnitine Palmitoyltransferase II and Impaired Mitochondrial Fuel Utilzation in Influenza-Associated Encephalopathy, Human Mutation, 29(5); 718-727 (2008)
  11. Iddamalgoda A, Le QT, Ito K, Tanaka K, Kojima H, Kido H.: Mast cell tryptase and photoaging: possible involvement in the degradation of extra cellular matrix and basement membrane proteins, Arch Dermatol Res. Oct 30, 300, 69-76 (2008)
  12. Hiroshi Yamada, Youichiro Higashi, Aoi Kousaka, Tadashi Enomoto and Hiroshi Kido: Diclofenac augments intracellular influenza virus replication, Options for the Control of Influenza VI, 633-635 (2008)
  13. Hiroshi Kido, Dai Mizuno, Tsunetomo Takaei, Maki Nishino, Wakako Shinahara: New effective nasal immunizations of influenza vaccine by a natural mucosal adjuvant from the lung and its synthetic compound, Options for the Control of Influenza VI, 616-618 (2008)
  14. Hiroshi Kido, Yuushi Okumura and Dong Ryong Kim: MSPL/TMPRSS13, Front Biosci., Jan 1;13:754-8 (2008)
  15. Mihiro Yano and Hiroshi Kido: The Molecular Chaperones in the Ubiquitin-Proteasome System, The Ubiquitin Proteasome System, Eds: Mario Di Napoli and Cezary Wojcik, pp. 207-225 (2007) Nova Science Publishers, Inc.
  16. Masamichi Kuwajima, Hiroaki Fujihara, Hiroyoshi Sei, Asako Umehara, Masako Sei, Tomi T. Tsuda, Akiko Sukeno, Tatsuya Okamoto, Yoichi Ueta, Toshio Dai and Hiroshi Kido: Reduced carnitine level suppresses hypothalamic orexin expression during weaning period., Endocr. J., 54(6): 911-925, (2007)
  17. Masaki Hayama, Yuushi Okumura, Etsuhisa Takahashi, Aki Shimabukuro, Mnabu Tamura, Noriaki Takeda, Takeshi Kubo and Hiroshi Kido: Idenifification and analysis of the promoter region of the type II transmembrane serine protease polyserase-1 and its transcript variants, Biol. Chem., 388(8), 853-858 (2007)
  18. Tsutomu Hamuro, Hiroshi Kido, Yujiro Asada, Kinta Hatakeyama, Yuushi Okumura, Youichi Kunori, Takashi Kamimura, Sadaaki Iwanaga and Shintaro Kamei: Tissue factor pathway inhibitor is highly susceptible to chymase-mediated proteolysis, FEBS J., 274,3065-3077 (2007)
  19. J. Aoki, T. Yasuno, H. Sugie, H. Kido, I. Nishino, Y. Shigematsu, M. Kanazawa, M. Takayanagi, M. Kumai, K. Endo, H. Kaneoka, M. Yamaguchi, T. Fukuda and T. Yamamoto: A Japanese Adult form of CPT II Deficiency Associated with a Homozygous F383Y. Mutation. Neurology, 69(8), 804-806 (2007)
  20. Mihiro Yano, Shinichi Nakamuta, Mayumi Shiota, Hiroshi Endo and Hiroshi Kido: Gatekeeper Role of 14-3-3 Protein in HIV-1 gp120-Mediated Apotosis of Human Endothelial Cells by Inactivation of Bad. AIDS, 21(8), 911-920 (2007)
  21. Hiroshi Kido, Yuushi Okumura, Hiroshi Yamada, Trong Quang Le and Mihiro Yano: Proteases essential for human influenza virus entry into the cells and their inhibitors as potential therapeutic agents. Curr. Pharma.Des., 13(3), 403-412 (2007)
  22. Dengfu Yao, Masamichi Kuwajima, Ye Chen, Mayumi Shiota, Yuushi Okumura, Hiroshi Yamada and Hiroshi
    Kido: Impaired long-chain fatty acid metabolism in mitochondria causes brain vascular invasion by a non-neurotropic epidemic influenza A virus in the newborn/suckling period: implications for influenza-associated encephalopathy. Mol. Cell. Biochem., 299, 85-92 (2007)
  23. Hiromi Yano, Naoki Muguruma, Susumu Ito, Eriko Aoyagi, Tetsuo Kimura, Yoshitaka Imoto, Jianxin Cao, Shohei Inoue, Shigeki Sano, Yoshimitsu Nagao, Hiroshi Kido:
    Fab fragment labeled with ICG-derivative for detecting digestive tract cancer. Photodiagnosis and Photodynamic Therapy 3, 177-183 (2006)
  24. Mihiro Yano, Shinichi Nakamuta, Xueji Wu. Yuushi Okumura and Hiroshi Kido: A novel function of 14-3-3 protein: 14-3-3 is a heat shock-related molecular chaperone that dissolves thermal-aggregated proteins. Mol. Biol. Cell, 17, 4769-4779 (2006)
  25. Yuushi Okumura, Masaki Hayama, Etsuhisa Takahashi, Miyoko Fujiuchi, Aki Shimabukuro, Mihiro Yano and Hiroshi Kido: Serase-1B, a new splice variant of polyserase –1/TMPRSS9, activates urokinase-type plasminogen activator and the proteolytic activation is negatively regulated by glycosamino-glycans. Biochem.J. 400, 551-561 (2006)
  26. Hiroshi Yamada, Quang Trong Le, Aoi Kousaka, Youichiro Higashi, Mari Tsukane and Hiroshi Kido: Sendai virus infection up-regulates trypsin I and matrix metalloproteinase-9, triggering viral multiplication and matrix degradation in rat lungs and lung L2 cells. Arch. Virol. 151, 2529-2537 (2006)
  27. K. Kunimi, M. Maegawa, M. Kamada, S. Yamamoto, T. Yasui, T. Matsuzaki, A. Kuwahara, H. Furumoto, Y. Ohmoto, H. Kido and M. Irahara. Myeloid-related protein-8/14 is associated with proinflammatory cytokines in cervical mucus. J. Reprod. Immunol., 71, 3-11 (2006)
  28. Trong Qunag Le, Miki Kawachi, Hiroshi Yamada, Mayumi Shiota,Yuushi Okumura and Hiroshi.
    Kido:Identification of Trypsin I as a candidate for influenza A virus and Sendai virus envelope glycoprotein processing protease in rat brain. Biol Chem., 387, 467-475 (2006)



(~2005) Major manuscripts in 154 manuscripts

  1. Masaki Kita, Yasuo nakamura, Yuushi Okumura, Satoshi D Ohdachi, Yuichi Oba, Michiyasu Yoshikuni, Hiroshi Kido, and Daisuke Uemura: Blarina toxin, a mammalian lethal venom from the short-tailed shrew Blarina brevicauda: Isolation and characterization. Proc. Natl. Acad. Sci. USA 101(20): 7542-7547 (2004)
  2. Cezary Wojcik, Mihiro Yano and George N. DeMartino: RNA interference of valosin-containing protein (VCP/p97) reveals multiple cellular roles linked to ubiquitin/proteasome-dependent proteolysis. J. Cell Sci. 117, 281-292 (2004)
  3. Xueji Wu, Mihiro Yano, Hiroyo Washida and Hiroshi Kido: The second metal-binding site of 70kDa heat-shock protein is essential for ADP-binding, ATP hydrolysis and ATP synthesis. Biochem. J. 378:793-799 (2004)
  4. Yuushi Okumura, Mai Nishikawa, Ping Cui, Mayumi Shiota, Yasuo Nakamura, Masataka Adachi, Kenichiro Kitamura, Kimio Tomita and Hiroshi Kido: Cloning and characterization of a novel transmembrane-type serine protease from rat kidney, a new member among the sodium channel activators. Biol. Chem. 384: 1483-1495 (2003)
  5. Masayuki Sato, Shuku Yoshida, Kemji Iida, Takanori Tomosawa, Hiroshi Kido and Makoto Yamashita: A novel influenza A virus activating enzyme from porcine lung: Purification and characterization. Biol. Chem. 384(2): 219-227 (2003)
  6. Yu-ichi Kamikubo, Yuushi Okumura and David J. Loskutof: Identification of the disulfide bonds in the recombinant somatomedine B domain of human vitronectin. J. Biol. Chem. 277(30): 27109-27119 (2002)
  7. T Towatari, M Ide, K Ohba, Y Chida, M Murakami, M Shiota, M Kawachi, H yamada and H Kido: Identification of ectopic anionic trypsin I in rat lungs potentiating pneumotropic virus infectivity and increased enzyme level after virus infection. Eur. J. Biochem. 269: 2613-2621 (2002)
  8. Bing Yang, Deng Fu yao, Masanobu Ohuchi, Mikiko Ide, Mihiro Yano, Yuushi Okumura and Hiroshi Kido: Ambroxol suppresses influenza-virus proliferation in the mouse airway by increasing antiviral factor levels. Eur. Res. J. 19(5): 952-958 (2002)
  9. Shuji Kondo, Shoji Kagami, Hiroshi Kido, Frabj Strutz, Gerhard A. Muller, and Yasuhiro Kuroda: Role of mast cell tryptase in renal intestinal fibrosis. J. Am. Soc. Nephrol. 12: 1668-1676 (2001)
  10. Meiko Murakami, Takae Towatari, Masanobu Ohuchi, Mayumi Shiota, Miyoko Akao, Yuushi Okumura, Marina A.A. Parry and Hiroshi Kido: Mini-plasmin found in the epithelial cells of bronchioles triggers infection by broad-spectrum influenza A viruses and Sendai virus. Eur. J. Biochem. 268(10): 2847-2855 (2001)
  11. Ye Chen, Maumi Shiota, Masanobu Ohuchi, Takae towatari, Junko Tashiro, Meiko Murakami, Mihiro Yano Bing Yang and Hiroshi Kido: mast cell tryptase from pig lungs triggers infection by pneumotropic Sendai and influenza A viruses. Eur. J. Biochem. 267(11): 3189-3197 (2000)
  12. M Inoue, M Isobe, T. Itoyama and H Kido: Structural analysis of esp-1 gene (PRSS 21). Biochem. Biophys. Res. Commun. 266(2): 564-568 (1999)
  13. Mihiro Yano, Sachie Mori and Hiroshi Kido: Intrinsic nucleoside diphosphate kinase-like activity is a novel function of the 20S proteasome. J. Biol. Chem. 274(48): 34375-34382 (1999)
  14. M Hara, A Matsumori, K Ono, H Kido, M.-W Hwang, T Miyamoto, A Iwasaki, M Okada, K Nakatani and S Sasayama: mast cell cause apoptosis of cardiomyocytes and proliferation of other intramyocardial mast cells in vitro. Circulation 100(13): 1443-1449 (1999)
  15. H Kido, M murakami, K Ohba, Y Chen and T Towatari: Cellular proteinases trigger the infectivity of the influenza A and Sendai viruses. Mol. Cells 9(3):235-244 (1999)
  16. Y Okumura, M Yano, M Murakami, T Towatari and H Kido: The extracellular processing of HIV-1 envelope glycoprotein gp160 by human plasmin. FEBS Lett. 442: 39-42 (1999)
  17. M Inoue, N Kanbe, M Kurosawa and H Kido: Cloning and tissue distribution of a novel serine protease esp-1 from human eosinophils. Biochem. Biophys. Res. Commun. 252(2): 307-312 (1998)
  18. M Hiromura, M Yano, H Mori, M Inoue and H Kido: Intrinsic ADP-ATP exchange activity is a novel function of the molecular chaperone, HSP70. J. Biol. Chem. 273(10): 5435-5438 (1998)
  19. A Nakano, F kishi, K Minami, H Wakabayashi, Y Nakaya and H Kido: Selective conversion of big endothelins to tracheal smooth muscle-constricting 31-amino acid length endothelins by chymase from human mast cells. J. Immunol. 159: 1987-1992 (1997)
  20. S Takada, H Kido, A Fukutomi, T Mori, and K Koike: Interaction of hepatitis B virus protein with a serine protease, tryptase TL2, as an inhibitor. Oncogene 9: 341-348 (1994)
  21. M Tashiro, M Takeda, S Tanaka, N Nishimura, M Takenaka and H Kido: Antibody against the carboxyl terminus of the F2 subunit of Sendai virus fusion glycoprotein inhibits proteolytic activation. Virology 194: 882-885 (1993)
  22. H Kido, K kamoshita, A Fukutomi, and N Katunuma: Processing protease for gp160 human immunodeficiency virus type I envelope glycoprotein precursor in human T4+ lymphocytes: Purification and characterization. J. Biol. Chem. 268(18): 13406-13413 (1993)
  23. M tashiro, Y Yokogoshi, K Tobita, J T. Seto, R Rott and H Kido: Tryptase Clara, an activating protease for Sendai virus in rat lungs, is involved in pneumopathogenicity. J. Virology 66(12): 7211-7216 (1992)
  24. H Kido, Y Yokogoshi, K Sakai, M Tashiro, Y Kishino, A Fukutomi, and N Katunuma: Isolation and characterization of a novel trypsin-like protease found in rat bronchiolar epithelial Clara cells: a possible activator of the viral fusion glycoprotein. J. Biol. Chem. 267(19): 13573-13579 (1992)