2001 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酸性糖蛋白 / S-ニトロソ化 / 酸化アルブミン / Cys残基 / ミオグロビン / 配位子結合性
Research Abstract	西は、α_1酸性糖蛋白(α_1-AGP)のマウス大動脈張力に対する効果を調べた。その結果、交感神経作用薬フェニレフリンで予め収縮させた標本に対し、α_1-AGPは0.3-3.0mg/mlの範囲で濃度依存性の弛緩を起こすことが示され、急性炎症期のα_1-AGP濃度上昇に伴い、血液循環の改善が行われていることが示唆された。前田は、α_1-AGPのS-ニトロソ(S-NO)化による新たな機能蛋白創製を試みた。ジチオスレイトールによる還元で生じたSH基とイソアミルナイトライトとの反応で調製されたS-NO-α_1-AGPを用いて、IL-8並びにFMLPに対する好中球の走化性に及ぼす抑制効果を調べた結果、α_1-AGPとの有意差は認められなかったものの、S-NO化が蛋白質機能の調節や新機能の付与など重要な翻訳後修飾の一つであることを既に見い出していることから、S-NO-α_1-AGPにおいても、α_1-AGPが有する血球循環改善や免疫調節の機能に加え、血流維持や組織保護など有用な新機能が付与されている可能性が示唆された。小田切は、酸化剤chloramine-Tにより調製された酸化アルブミン及びCys-34をSerに換えたアルプミン変異体の特性を、それぞれ未処理ヒトアルブミン及び野生型アルブミンと比較した。その結果、酸化の初期段階でCys残基が酸化されることが示され、さらに、Cys-34のみならずMet残基などの酸化も体内動態に影響を及ぼすことが示唆された。宇野は、大腸菌による大量発現系を用いて調製された酸素結合性の各種ミオグロビン変異体の配位子結合性及び分子構造について検討した。その結果、配位子の入口部分に酸性残基が存在すると配位子結合性が低下することが示された。また、ヘム鉄に軸配位している近位ヒスチジンを含めた四重変異をほどこすことにより、近位側に新しい配位子結合性を有するヘムポケットの構築に成功した。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] Tokutomi, Y., et al.: "The properties of ryanodine-sensitive Ca^<2+> release in mouse gastric smooth muscle cells"Br. J. Pharmacol.. 133. 125-137 (2001)
[Publications] Tokutomi, Y., et al.: "消化管自律運動の遺伝子基盤と病態"日本薬理学雑誌. 119・4(in press). (2002)
[Publications] Okuda, S., et al.: "Noradrenaline receptor-mediated potentiation of caffeine-induced Ca^<2+>-activated K^+ currents in bovine ciliary muscle cells"Current Eye Research. (in press). (2002)
[Publications] Tokutomi, Y., et al.: "The Effects of α_1-acid glycoprotein on isometric tension of mouse aorte"The Japanese Journal of Pharmacology. 88 supple1. 255P (2002)
[Publications] T.Okamoto, T.Akaike, T.Sawa, Y.Miyamoto, Albert van der Vliet, H.Maeda: "Activation of matrix metalloproteinases by peroxynitrite-induced protein S-glutathiolation via disulfide S-oxide formation"J. Biol. Chem. 276. 29596-29602 (2001)
[Publications] J.Wu, T.Akaike, K.Hayashida, T.Okamoto, A.Okuyama: "Enhanced vascular permeability in solid tumor involving peroxynitrite and matrix metalloproteinase"Jpn. J. Cancer Res.. 92. 439-451 (2001)
[Publications] T.Akaike: "Role of free radicals in viral pathogenesis and mutation"Rev. Med. Virol.. 11. 87-101 (2001)
[Publications] T.Sakai, M.Otagiri et al.: "Interaction Mechanism between Indoxyl Sulfate, a Typical Uremic Toxin Bound to SiteII, and Ligands Bound to SiteI of Human Serum Albumin"Pharm. Res.. 18(4). 520-524 (2001)
[Publications] J.S.Zhang, M.Otagiri et al.: "Stability of a Cisplatin-Chondroitin Sulfate A Complex in Plasma and Kidney in Terms of Protein Binding"Biol. Pharm. Bull.. 24(8). 970-972 (2001)
[Publications] M.Anraku, M.Otagiri et al.: "Effect of Oxidative Stress on the Structure and Function of Human Serum Albumin"Pharm. Res.. 18(5). 269-274 (2001)
[Publications] H.Watanabe, M.Otagiri et al.: "Conformational Stability and Warfarin-binding Properties of Human Serum Albumin Studied by Recombinant Mutants"Biochem. J.. 357. 666-671 (2001)
[Publications] K.Shimoishi, M.Otagiri: "FK506(tacrolimus) Increases α_1-acid Glycoprotein Expression in Liver and Primary Cultured Hepatocytes"Eu. J. Pharmacol.. 420. 91-95 (2001)
[Publications] T.Komori, M.Otagiri et al.: "Clarithromycin Up-regulates α_1-Acid Glycoprotein. Expression in Liver and Primary cultured Hepatocytes"Biochem. Pharmacol. 62. 1391-1397 (2001)
[Publications] H.Watanabe, M.Otagiri et al.: "In Vitro and In Vivo Properties of Recombinant Human Serum Albumin from Pichia Pastoris Purified by a Method of Short Processing Time"Pharm. Res.. 18(12). 1775-1781 (2001)
[Publications] K.Funato, T.Imai, K.Nakashima, M.Otagiri: "High-performance liquid chromatography with chemiluminescence detection of penbutolol and its hydroxylated metabolite in rat plasma"J Chromatogr B Biomed Sci Appl.. 757(2). 229-235 (2001)
[Publications] H.Imamura, M.Otagiri et al.: "Stereoselective Protein Binding of Alprenolol in the Renal Diseased State"Chirality. 14(in press). (2002)
[Publications] T.Deguchi, S.Ohtsuki, M.Otagiri et al.: "Major Role of Organic Anion Transporter 3 in the Transport of Indoxil Sulfate in the Kidney"Kidney. Int.. (in press). (2002)
[Publications] V.T.G.Chuang, U.K-Hansen, M.Otagiri: "Pharmaceutical Strategies Utilizing Recombinant Human Serum Albumin (Review)"Pharm. Res.. (in press). (2002)
[Publications] Tadayuki Uno, Akihiro Yukinari, Yoshikazu Tomisugi, Yoshinobu Ishikawa, Ryu Makino, James A. Brannigan, Anthony J. Wilkinson: "Cysteine Thiolate Coordination in the Ferrous CO Complex of an Engineered Cytochrome b562"J. Am. Chem. Soc.. 123. 2458-2459 (2001)
[Publications] Yoshinobu Ishikawa, Takeshi Yamashita, Yoshikazu Tomisugi, Tadayuki Uno: "Interaction of porphyrins bearing peripheral cationic heterocycles with G-quadruplex DNA"Nucleic Acids Res.. Suppl.1. 107-108 (2001)
[Publications] Naoki Yamakawa, Yoshinobu Ishikawa, Tadayuki Uno: "Solution Properties and Photonuclease Activity of Cationic Bis-porphyrins Linked with a Series of Aliphatic Diamines"Chem. Pharm. Bull.. 49. 1531-1540 (2001)
[Publications] Yoshinobu Ishikawa, Naoki Yamakawa, Tadayuki Uno: "Potent DNA Photocleavage by Zinc(II) Complexes of Cationic Bis-porphyrins Linked with Aliphatic Diamine"Bioorg. Med. Chem.. 10(in press). (2001)
[Publications] Tadayuki Uno, Rikiharu Sakamoto, Yoshikazu Tomisugi, Yosbinobu Ishikawa, John S.Olson, Anthony J.Wilkinson: "Inversion of Axial Coordination in Myoglobin to Create a "Proximal" Ligand Binding Pocket"J. Am. Chem. Soc.. (submitted, in press). (2002)

2001 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, Y., et al.: "The properties of ryanodine-sensitive Ca^<2+> release in mouse gastric smooth muscle cells"Br. J. Pharmacol.. 133. 125-137 (2001)

[Publications] Tokutomi, Y., et al.: "消化管自律運動の遺伝子基盤と病態"日本薬理学雑誌. 119・4(in press). (2002)

[Publications] Okuda, S., et al.: "Noradrenaline receptor-mediated potentiation of caffeine-induced Ca^<2+>-activated K^+ currents in bovine ciliary muscle cells"Current Eye Research. (in press). (2002)

[Publications] Tokutomi, Y., et al.: "The Effects of α_1-acid glycoprotein on isometric tension of mouse aorte"The Japanese Journal of Pharmacology. 88 supple1. 255P (2002)

[Publications] T.Okamoto, T.Akaike, T.Sawa, Y.Miyamoto, Albert van der Vliet, H.Maeda: "Activation of matrix metalloproteinases by peroxynitrite-induced protein S-glutathiolation via disulfide S-oxide formation"J. Biol. Chem. 276. 29596-29602 (2001)

[Publications] J.Wu, T.Akaike, K.Hayashida, T.Okamoto, A.Okuyama: "Enhanced vascular permeability in solid tumor involving peroxynitrite and matrix metalloproteinase"Jpn. J. Cancer Res.. 92. 439-451 (2001)

[Publications] T.Akaike: "Role of free radicals in viral pathogenesis and mutation"Rev. Med. Virol.. 11. 87-101 (2001)

[Publications] T.Sakai, M.Otagiri et al.: "Interaction Mechanism between Indoxyl Sulfate, a Typical Uremic Toxin Bound to SiteII, and Ligands Bound to SiteI of Human Serum Albumin"Pharm. Res.. 18(4). 520-524 (2001)

[Publications] J.S.Zhang, M.Otagiri et al.: "Stability of a Cisplatin-Chondroitin Sulfate A Complex in Plasma and Kidney in Terms of Protein Binding"Biol. Pharm. Bull.. 24(8). 970-972 (2001)

[Publications] M.Anraku, M.Otagiri et al.: "Effect of Oxidative Stress on the Structure and Function of Human Serum Albumin"Pharm. Res.. 18(5). 269-274 (2001)

[Publications] H.Watanabe, M.Otagiri et al.: "Conformational Stability and Warfarin-binding Properties of Human Serum Albumin Studied by Recombinant Mutants"Biochem. J.. 357. 666-671 (2001)

[Publications] K.Shimoishi, M.Otagiri: "FK506(tacrolimus) Increases α_1-acid Glycoprotein Expression in Liver and Primary Cultured Hepatocytes"Eu. J. Pharmacol.. 420. 91-95 (2001)

[Publications] T.Komori, M.Otagiri et al.: "Clarithromycin Up-regulates α_1-Acid Glycoprotein. Expression in Liver and Primary cultured Hepatocytes"Biochem. Pharmacol. 62. 1391-1397 (2001)

[Publications] H.Watanabe, M.Otagiri et al.: "In Vitro and In Vivo Properties of Recombinant Human Serum Albumin from Pichia Pastoris Purified by a Method of Short Processing Time"Pharm. Res.. 18(12). 1775-1781 (2001)

[Publications] K.Funato, T.Imai, K.Nakashima, M.Otagiri: "High-performance liquid chromatography with chemiluminescence detection of penbutolol and its hydroxylated metabolite in rat plasma"J Chromatogr B Biomed Sci Appl.. 757(2). 229-235 (2001)

[Publications] H.Imamura, M.Otagiri et al.: "Stereoselective Protein Binding of Alprenolol in the Renal Diseased State"Chirality. 14(in press). (2002)

[Publications] T.Deguchi, S.Ohtsuki, M.Otagiri et al.: "Major Role of Organic Anion Transporter 3 in the Transport of Indoxil Sulfate in the Kidney"Kidney. Int.. (in press). (2002)

[Publications] V.T.G.Chuang, U.K-Hansen, M.Otagiri: "Pharmaceutical Strategies Utilizing Recombinant Human Serum Albumin (Review)"Pharm. Res.. (in press). (2002)

[Publications] Tadayuki Uno, Akihiro Yukinari, Yoshikazu Tomisugi, Yoshinobu Ishikawa, Ryu Makino, James A. Brannigan, Anthony J. Wilkinson: "Cysteine Thiolate Coordination in the Ferrous CO Complex of an Engineered Cytochrome b562"J. Am. Chem. Soc.. 123. 2458-2459 (2001)

[Publications] Yoshinobu Ishikawa, Takeshi Yamashita, Yoshikazu Tomisugi, Tadayuki Uno: "Interaction of porphyrins bearing peripheral cationic heterocycles with G-quadruplex DNA"Nucleic Acids Res.. Suppl.1. 107-108 (2001)

[Publications] Naoki Yamakawa, Yoshinobu Ishikawa, Tadayuki Uno: "Solution Properties and Photonuclease Activity of Cationic Bis-porphyrins Linked with a Series of Aliphatic Diamines"Chem. Pharm. Bull.. 49. 1531-1540 (2001)

[Publications] Yoshinobu Ishikawa, Naoki Yamakawa, Tadayuki Uno: "Potent DNA Photocleavage by Zinc(II) Complexes of Cationic Bis-porphyrins Linked with Aliphatic Diamine"Bioorg. Med. Chem.. 10(in press). (2001)

[Publications] Tadayuki Uno, Rikiharu Sakamoto, Yoshikazu Tomisugi, Yosbinobu Ishikawa, John S.Olson, Anthony J.Wilkinson: "Inversion of Axial Coordination in Myoglobin to Create a "Proximal" Ligand Binding Pocket"J. Am. Chem. Soc.. (submitted, in press). (2002)

西勝英熊本大学, 医学部, 教授 (00040220)