2000 Fiscal Year Annual Research Report

血液蛋白成分(α-1酸性糖蛋白)の有効利用と血液代替物質の開発研究

Research Project

Project/Area Number	11794016
Research Institution	Kumamoto University
Principal Investigator	西勝英熊本大学, 医学部, 教授 (00040220)
Co-Investigator(Kenkyū-buntansha)	宇野公之熊本大学, 薬学部, 教授 (00183020) 小田切優樹熊本大学, 薬学部, 教授 (80120145) 前田浩熊本大学, 医学部, 教授 (90004613)
Keywords	α-1酸性糖タンパク / 細隙通過促進作用 / 微細循環改善作用 / 白血球走化性抑制作用 / 炎症 / 変異アルブミン / ミオグロビン変異体 / シトクロムb562変異体
Research Abstract	西らは、α-1酸性糖タンパクの赤血球の細隙通過促進作用について検討を行った。生理食塩水に10%ヒト赤血球浮遊液を調整し、100μlの浮遊液の直径3μ、5μの細隙通過時間を測定し、α-1酸性糖タンパク添加時の通過時間との比較をおこなったところ、α-1酸性糖タンパク10μg/ml〜100μg/mlの添加により、有意に細隙通過時間の短縮をみとめ、α-1酸性糖タンパクの微細循環改善作用の一端を明らかにした。前田らは、a1-酸性蛋白の抗炎症作用を、白血球走化性の抑制作用に焦点をあて解析した。a1-酸性蛋白は、モルモット好中球のヒトレコンビナントinterlerkin-8およびN-formyl-methionyl-leucyl-phenylalanine(fMLP)による走化性誘導を強く抑制した。この結果は、a1-酸性蛋白が炎症局所における各種ケモカインや走化性因子による炎症誘導に対して抑制的に作用する可能性を示唆する。小田切らは、アルブミン分子上の、199-,439-,525-LysをAla残基に置換した変異体を作製し、その構造と機能について調べたところ、変異に伴う構造、機能変化は認められないものの、これら変異アルブミンの生体内半減期はnativeアルブミンと変わらないか、むしろ短縮する傾向を示した。宇野らは、大腸菌による大量発現系を用いて、酸素結合性を有するミオグロビン変異体、及びシトクロムb562変異体を調製した。これらの配位子結合性を分光学的に調べ、また共鳴ラマンスペクトルの測定を通して分子構造の観点から検討を加えた。その結果、ミオグロビンの近位水素結合網や配位子の入口部分における水素結合網の崩壊により、配位子結合性が向上することがわかった。また、シトクロムb562の多重変異により配位子結合性ポケットの構築に成功した。

Research Products

(31 results)

All Other

All Publications (31 results)

[Publications] Tokutomi N.Nishi K.: "The Effects of Mexiletine on the Activity of Neuronal Voltage-gated Na^+ channels"Jpn.Pharmacol & Therapeutics. 28. 133-141 (2000)
[Publications] Zhong-Fang Lai,Yu-Zhen Chen,Yasuharu Nishimura,and Katsuhide Nishi: "An Amiloride-Sensitive and Voltage-Dependent Na^+ Channel in an HLA-DR-Restricted Human T Cell Clone^1"The Journal of Immunology. 165. 83-90 (2000)
[Publications] Zhong-Fang Lai & Katsuhide Nishi: "Enhancement of Intracellular Cl^- Concentrations Induced by Extracellular ATP in Guinea Pig Ventricurar Muscle"Jpn.J.Phamacol. 84. 438-448 (2000)
[Publications] Nishi K.,Nakaishi Y.and Maeda H.: "Potential physiological role of alpha-1 acid glycoprotein to facilitate the assage of erythrocytes in microcirculation"International Symposium on Serum Albumin & α1-Acid Glycoprotein. 67-68 (2000)
[Publications] Tokutomi,Y.,Tokutomi,N.and Nishi K.: "The properties of ryanodine-sensitive Ca^<2+> release in mouse gastric smooth muscle cells."British Journal of Pharmacology. (in press). (2000)
[Publications] Y.Miyamoto,T.Akaike,M.Samiul Alam,K.Inoue,T.Hamamoto,N.Ikebe,J.Yoshitake,T.Okamoto and H.Maeda: "Novel functions of human _1-protease inhibitor after S-nitrosylation : inhibition of cysteine protease and antibacterial activity."Biochem. Biophys. Res.Commun.. 267. 918-923 (2000)
[Publications] T.Akaike,S.Fujii,A.Kato,J.Yoshitake,Y.Miyamoto,T.Sawa,S.Okamoto,M.Suga,M.Asakawa,Y.Nagai and H.Maeda: "Viral mutation accelerated by nitric oxide production during infection in vivo."FASEB J.. 14. 1447-1454 (2000)
[Publications] T.Sawa,J.Wu,T.Akaike and H.Maeda: "Tumor-targeting chemotherapy by a xanthine oxidase-polymer conjugate that generates oxygen-free radicals in tumor tissue."Cancer Res.. 60. 666-671 (2000)
[Publications] H.Maeda,J.Wu,T.Sawa,Y.Matsumura and K.Hori: "Tumor vascular permeability and the EPR effect in macromolecular therapeutics."J.Controlled Release. 65. 271-284 (2000)
[Publications] Y.Miyamoto,T.Akaike and H.Maeda: "S-Nitrosylated human _1-protease inhibitor."Biochim. Biophys. Acta. 1477. 90-97 (2000)
[Publications] H.Kuwahara,Y.Miyamoto,T.Akaike,T.Kubota,T.Sawa,S.Okamoto and H.Maeda: "Helicobacter pylori urease suppresses bactericidal activity of peroxynitrite viacarbon dioxide production."Infect.Immun.. 63. 4378-4383 (2000)
[Publications] T.Sawa,T.Akaike and H.Maeda: "Tyrosine nitration by peroxynitrite formed from nitric oxide and superoxide generated by xanthine oxidase."J.Biol.Chem.. 275. 32467-32474 (2000)
[Publications] E.Akizuki,T.Akaike,S.Okamoto,S.Fujii,Y.Yamaguchi,M.Ogawa and H.Maeda: "Role of nitric oxide and superoxide in acute cardiac allograft rejection in rats."Proc.Soc.Exp.Biol.Med.. 225. 151-159 (2000)
[Publications] N.Ikebe,T.Akaike,Y.Miyamoto,K.Hayashida,J.Yoshitake,M.Ogawa and H.Maeda: "Protective effect of S-nitroso-_1-protease inhibitor on hepatic ischemia-reperfusion injury."J.Pharmacol.Exp.Ther.. 295. 904-911 (2000)
[Publications] T.Akaike,and H.Maeda: "Nitric oxide and virus infection."Immunology. 101. 300-308 (2000)
[Publications] H.Watanabe,S.Tanase,M.Otagiri et al.: "Role of Arg-410 and Tyr-411 in Human Serum Albumin for Ligand Binding and Esterase-like Activity."Biochem.J.. 349. 813-819 (2000)
[Publications] J.Zhang,T.Imai,M.Otagiri.: "Effects of a Cisplatin-Chondroitin Sulfate A Complex on Reducing the Nephrotoxicity of Cisplatin."Arch.Toxicol. 74(6). 300-307 (2000)
[Publications] N.Kurata,S.Matsushita,M.Otagiri et al.: "Characterization of a Binding Site of UCN-01, a Novel Anticancer Drug on _1-Acid Glycoprotein."Biol.Pharm.Bull. 23(7). 893-895 (2000)
[Publications] K.Yamasaki,H.M.Rahman,M.Otagiri et al.: "Circular Dichroism Simulation shows a Site-II-Site-I Displacement of Human Serum Albumin-bound Diclofenac by Ibuprofen."AAPS Pharm.Sci.Tech.,. 1(2)(article 12). (2000)
[Publications] Y.Tsutsumi,T.Maruyama,M.Otagiri et al.: "Decreased Bilirubin-binding Capacity in Uremic Serum Caused by an Acumulation of Furan Dicarboxylic Acid."Nephron. 85(1). 60-64 (2000)
[Publications] C.Kawasaki,R.Nishi,M.Otagiri et al.: "Chacoal Hemoperfusion in the Treatment of Phenytoin Overdose."Am.J.Kidney Dis.. 35(2). 323-326 (2000)
[Publications] V.C.T.Gian,H.Nakayama,M.Otagiri et al.: "Helix 6 of Subdomain IIIA of Human Serum Albumin is the Region Primarily Photolabeled by Ketoprofen, an Arylpropionic Acid NSAID Containing a Benzophenone Moiety."Biochim. Biophys. Acta,. 1434. 18-30 (1999)
[Publications] K.Yamasaki,T.Maruyama,M.Otagiri et al.: "Interactive Binding to the Two Principal Ligand Binding Sites of Human Serum Albumin : Effect of the Neutral-to-Base Transition."Biochim. Biophys. Acta,. 1432. 313-323 (1999)
[Publications] T.Sakai,S.Fujiyama,M.Otagiri et al.: "Stereoselective Serum Protein Binding of Ketoprofen in Liver Diseases."Enantiome. 4(5). 477-482 (1999)
[Publications] Y.Tsutsumi,T.Maruyama,M.Otagiri et al.: "Interaction between Two Dicarboxylate Endogenous Substances, Bilirubin and Uremic Toxin, 3-Carboxy-4-methyl-5-propyl-2-furanpropanoic Acid, on Human Serum Albumin."Pharm.Res.. 16(6). 916-923 (1999)
[Publications] T.Imai,M.Taketani,M.Otagiri et al.: "In Vitro Identification of the Human Cytochrome P-450 Enzymes Involved in the N-Demethylation of Azelastine."Drug.Metab.Dispos.. 27(8). 942-946 (1999)
[Publications] T.Imai,M.Sakai,M.Otagiri et al: "In Vitro and In Vivo Evaluation of the Enhancing Activity of Glycyrrlizin on the Intestinal Absorption of Drugs."Pharm.Res.. 16(1). 80-86 (1999)
[Publications] Keitarou Suzuki,Fumiko Shono,Hirofumi Kai,Tadayuki Uno,and Masaru Uyeda: "Inhibition of Topoisomerases by Fatty Acids"J.Enzyme Inhibition. 15. 357-366 (2000)
[Publications] Yoshinobu Ishikawa,Aya Yamashita,and Tadayuki Uno: "Efficient photocleavage of DNA by cationic porphyrin-acridine hybrids with the effective length of diamino alky linkage"Chem.Pharm.Bull.. 49(in press). (2001)
[Publications] Tadayuki Uno,Akihiro Yukinari,Yoko Moriyama,Yoshinobu Ishikawa,Yoshikazu Tomisugi,James A.Brannigan,and Anthony J. Wilkinson: "Engineering a Ligand Binding Heme Pocket into a Four Helix Bundle Protein Cytochrome b562"J.Am.Chem.Soc.. 123. 512-513 (2001)
[Publications] Tadayuki Uno,Akihiro Yukinari,Yoshikazu Tomisugi,Yoshinobu Ishikawa,Ryu Makino,James A.Brannigan,and Anthoy J.Wilkinson: "Cysteine Thiolate Coordination in the Ferrous CO Complex of an Engineered Cytochrome b562"J.Am.Chem.Soc.. 123(in press). (2001)

2000 Fiscal Year Annual Research Report

血液蛋白成分(α-1酸性糖蛋白)の有効利用と血液代替物質の開発研究

Principal Investigator

西 勝英 熊本大学, 医学部, 教授 (00040220)

Research Products

[Publications] Tokutomi N.Nishi K.: "The Effects of Mexiletine on the Activity of Neuronal Voltage-gated Na^+ channels"Jpn.Pharmacol & Therapeutics. 28. 133-141 (2000)

[Publications] Zhong-Fang Lai,Yu-Zhen Chen,Yasuharu Nishimura,and Katsuhide Nishi: "An Amiloride-Sensitive and Voltage-Dependent Na^+ Channel in an HLA-DR-Restricted Human T Cell Clone^1"The Journal of Immunology. 165. 83-90 (2000)

[Publications] Zhong-Fang Lai & Katsuhide Nishi: "Enhancement of Intracellular Cl^- Concentrations Induced by Extracellular ATP in Guinea Pig Ventricurar Muscle"Jpn.J.Phamacol. 84. 438-448 (2000)

[Publications] Nishi K.,Nakaishi Y.and Maeda H.: "Potential physiological role of alpha-1 acid glycoprotein to facilitate the assage of erythrocytes in microcirculation"International Symposium on Serum Albumin & α1-Acid Glycoprotein. 67-68 (2000)

[Publications] Tokutomi,Y.,Tokutomi,N.and Nishi K.: "The properties of ryanodine-sensitive Ca^<2+> release in mouse gastric smooth muscle cells."British Journal of Pharmacology. (in press). (2000)

[Publications] Y.Miyamoto,T.Akaike,M.Samiul Alam,K.Inoue,T.Hamamoto,N.Ikebe,J.Yoshitake,T.Okamoto and H.Maeda: "Novel functions of human _1-protease inhibitor after S-nitrosylation : inhibition of cysteine protease and antibacterial activity."Biochem. Biophys. Res.Commun.. 267. 918-923 (2000)

[Publications] T.Akaike,S.Fujii,A.Kato,J.Yoshitake,Y.Miyamoto,T.Sawa,S.Okamoto,M.Suga,M.Asakawa,Y.Nagai and H.Maeda: "Viral mutation accelerated by nitric oxide production during infection in vivo."FASEB J.. 14. 1447-1454 (2000)

[Publications] T.Sawa,J.Wu,T.Akaike and H.Maeda: "Tumor-targeting chemotherapy by a xanthine oxidase-polymer conjugate that generates oxygen-free radicals in tumor tissue."Cancer Res.. 60. 666-671 (2000)

[Publications] H.Maeda,J.Wu,T.Sawa,Y.Matsumura and K.Hori: "Tumor vascular permeability and the EPR effect in macromolecular therapeutics."J.Controlled Release. 65. 271-284 (2000)

[Publications] Y.Miyamoto,T.Akaike and H.Maeda: "S-Nitrosylated human _1-protease inhibitor."Biochim. Biophys. Acta. 1477. 90-97 (2000)

[Publications] H.Kuwahara,Y.Miyamoto,T.Akaike,T.Kubota,T.Sawa,S.Okamoto and H.Maeda: "Helicobacter pylori urease suppresses bactericidal activity of peroxynitrite viacarbon dioxide production."Infect.Immun.. 63. 4378-4383 (2000)

[Publications] T.Sawa,T.Akaike and H.Maeda: "Tyrosine nitration by peroxynitrite formed from nitric oxide and superoxide generated by xanthine oxidase."J.Biol.Chem.. 275. 32467-32474 (2000)

[Publications] E.Akizuki,T.Akaike,S.Okamoto,S.Fujii,Y.Yamaguchi,M.Ogawa and H.Maeda: "Role of nitric oxide and superoxide in acute cardiac allograft rejection in rats."Proc.Soc.Exp.Biol.Med.. 225. 151-159 (2000)

[Publications] N.Ikebe,T.Akaike,Y.Miyamoto,K.Hayashida,J.Yoshitake,M.Ogawa and H.Maeda: "Protective effect of S-nitroso-_1-protease inhibitor on hepatic ischemia-reperfusion injury."J.Pharmacol.Exp.Ther.. 295. 904-911 (2000)

[Publications] T.Akaike,and H.Maeda: "Nitric oxide and virus infection."Immunology. 101. 300-308 (2000)

[Publications] H.Watanabe,S.Tanase,M.Otagiri et al.: "Role of Arg-410 and Tyr-411 in Human Serum Albumin for Ligand Binding and Esterase-like Activity."Biochem.J.. 349. 813-819 (2000)

[Publications] J.Zhang,T.Imai,M.Otagiri.: "Effects of a Cisplatin-Chondroitin Sulfate A Complex on Reducing the Nephrotoxicity of Cisplatin."Arch.Toxicol. 74(6). 300-307 (2000)

[Publications] N.Kurata,S.Matsushita,M.Otagiri et al.: "Characterization of a Binding Site of UCN-01, a Novel Anticancer Drug on _1-Acid Glycoprotein."Biol.Pharm.Bull. 23(7). 893-895 (2000)

[Publications] K.Yamasaki,H.M.Rahman,M.Otagiri et al.: "Circular Dichroism Simulation shows a Site-II-Site-I Displacement of Human Serum Albumin-bound Diclofenac by Ibuprofen."AAPS Pharm.Sci.Tech.,. 1(2)(article 12). (2000)

[Publications] Y.Tsutsumi,T.Maruyama,M.Otagiri et al.: "Decreased Bilirubin-binding Capacity in Uremic Serum Caused by an Acumulation of Furan Dicarboxylic Acid."Nephron. 85(1). 60-64 (2000)

[Publications] C.Kawasaki,R.Nishi,M.Otagiri et al.: "Chacoal Hemoperfusion in the Treatment of Phenytoin Overdose."Am.J.Kidney Dis.. 35(2). 323-326 (2000)

[Publications] V.C.T.Gian,H.Nakayama,M.Otagiri et al.: "Helix 6 of Subdomain IIIA of Human Serum Albumin is the Region Primarily Photolabeled by Ketoprofen, an Arylpropionic Acid NSAID Containing a Benzophenone Moiety."Biochim. Biophys. Acta,. 1434. 18-30 (1999)

[Publications] K.Yamasaki,T.Maruyama,M.Otagiri et al.: "Interactive Binding to the Two Principal Ligand Binding Sites of Human Serum Albumin : Effect of the Neutral-to-Base Transition."Biochim. Biophys. Acta,. 1432. 313-323 (1999)

[Publications] T.Sakai,S.Fujiyama,M.Otagiri et al.: "Stereoselective Serum Protein Binding of Ketoprofen in Liver Diseases."Enantiome. 4(5). 477-482 (1999)

[Publications] Y.Tsutsumi,T.Maruyama,M.Otagiri et al.: "Interaction between Two Dicarboxylate Endogenous Substances, Bilirubin and Uremic Toxin, 3-Carboxy-4-methyl-5-propyl-2-furanpropanoic Acid, on Human Serum Albumin."Pharm.Res.. 16(6). 916-923 (1999)

[Publications] T.Imai,M.Taketani,M.Otagiri et al.: "In Vitro Identification of the Human Cytochrome P-450 Enzymes Involved in the N-Demethylation of Azelastine."Drug.Metab.Dispos.. 27(8). 942-946 (1999)

[Publications] T.Imai,M.Sakai,M.Otagiri et al: "In Vitro and In Vivo Evaluation of the Enhancing Activity of Glycyrrlizin on the Intestinal Absorption of Drugs."Pharm.Res.. 16(1). 80-86 (1999)

[Publications] Keitarou Suzuki,Fumiko Shono,Hirofumi Kai,Tadayuki Uno,and Masaru Uyeda: "Inhibition of Topoisomerases by Fatty Acids"J.Enzyme Inhibition. 15. 357-366 (2000)

[Publications] Yoshinobu Ishikawa,Aya Yamashita,and Tadayuki Uno: "Efficient photocleavage of DNA by cationic porphyrin-acridine hybrids with the effective length of diamino alky linkage"Chem.Pharm.Bull.. 49(in press). (2001)

[Publications] Tadayuki Uno,Akihiro Yukinari,Yoko Moriyama,Yoshinobu Ishikawa,Yoshikazu Tomisugi,James A.Brannigan,and Anthony J. Wilkinson: "Engineering a Ligand Binding Heme Pocket into a Four Helix Bundle Protein Cytochrome b562"J.Am.Chem.Soc.. 123. 512-513 (2001)

[Publications] Tadayuki Uno,Akihiro Yukinari,Yoshikazu Tomisugi,Yoshinobu Ishikawa,Ryu Makino,James A.Brannigan,and Anthoy J.Wilkinson: "Cysteine Thiolate Coordination in the Ferrous CO Complex of an Engineered Cytochrome b562"J.Am.Chem.Soc.. 123(in press). (2001)

西勝英熊本大学, 医学部, 教授 (00040220)