1997 Fiscal Year Annual Research Report

ジーンターゲテイングによる分泌型プロスタグランジンD合成酵素の機能解析

Research Project

Project/Area Number	07457033
Research Institution	Osaka Bioscience Institute
Principal Investigator	裏出良博 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (10201360)
Co-Investigator(Kenkyū-buntansha)	金岡禧秀 (財)大阪バイオサイエンス研究所, 第2研究部, 研究員 (40271514) 松村人志 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (50173886) 早石修 (財)大阪バイオサイエンス研究所, 第2研究部, 所長、部長 (40025507)
Keywords	睡眠 / 覚醒 / プロスタグランジンD2 / プロスタグランジンD合成酵素 / 遺伝子工学 / ノックアウトマウス / リポカリン / ベータートレース
Research Abstract	ラットおよびヒト脳における主要プロスタグランジン(PG)であるPGD2は、強力な睡眠誘発作用を示し、痛覚や臭覚の制御などの多彩な中枢作用を示す。中枢神経系におけるPGD2の生合成反応を触媒し生理的睡眠調節の鍵を握る酵素と考えられているリポカリン型PGD合成酵素は、そのcDNAクローニングの結果、各種の分泌蛋白質で構成されるリポカリンファミリーに属する唯一の酵素であることが判明した。本酵素は脳・脊髄などの中枢神経系や雄性生殖器に分布し、bトレース蛋白質として脳脊髄液や精液中に分泌される。本酵素は疎水性リガンドの輸送蛋白質が進化の過程で活性中心アミノ酸(Cys65)を獲得して酵素に進化したと考えられるが、他のリポカリン蛋白質と同様、レチノイドや甲状腺ホルモン・胆汁色素などの疎水性物質を特異的に結合する特性を保持していた。そこで、マウスの本酵素遺伝子を単離し、本酵素遺伝子を欠損したノックアウトマウスを作製した。その結果、遺伝子欠損ホモ・マウスが得られ、本酵素の遺伝子欠損は致死性を伴わないことが判明した。又、得られた遺伝子欠損マウスを用いて、その遺伝子欠損が繁殖率の異常や痛覚反応の異常をもたらすことを証明した。さらに、マウスの本酵素cDNAを大腸菌内で発現させた遺伝子組換え型酵素を用いて、世界で初めてリポカリン型PGD合成酵素の結晶化に成功した。

Research Products
(24 results)

All Other

All Publications (24 results)

[Publications] Yoshihiro Urada: "Lipid Mediators Cell Signalling" J.Neurosci.257-273. 12. 257-273 (1995)
[Publications] Makoto Murakami: "Interleukin-4 suppresses c-Kit ligand-induced expression of cytosolic phospholipase A2 and prostaglandin endoperoxide synthase-2,and their roles in separate pathways of eicosanoid synthesis in mouse bone marrow-derived mast sells." Proc.Natl.Acad.Sci.USA. 92. 6107-6111 (1995)
[Publications] Bettina Blodorn: "he major site of mRNA expression for the b-trace protein (prostaglandin D synthase) in human brain." Neurosci.Lett.209. 117-120 (1996)
[Publications] Hiroshi Oda: "Quantitative sandwich immunosorbent assay of human secretory prostaglandin D synthase (b-trace)." Proc.Japan Acad.72. 108-111 (1996)
[Publications] Carsten T.Beuckmann: "Lipocalin-type prostaglandin D synthase (b-trace) is located in pigmented epithelial cells of rat retina and accumulates within interphotoreceptor matrix." J.Neurosci.16. 6119-6124 (1996)
[Publications] Yoshihiro Urade: "Molecular mechanism of sleep regulation by prostaglandin D2. J." Lipid Mediators Cell Signalling. 14. 71-82 (1996)
[Publications] Dimitrios N.Melegos: "Immunofluorometric assay of prostaglandin D synthase (b-trace) and preliminary clinical applications." Clin.Chem.42. 1984-1991 (1996)
[Publications] Hiroyuki Toh: "IGlutathione-independent prostaglandin D synthase as a lead molecule for designing new functional proteins." Protein Engineering. 9. 1067-1082 (1996)
[Publications] Tetsumori Yamashima: "IProstaglandin D synthase (b-trace) in human arachnoid and meningioma cells : roles as a cell marker or in CSF absorption,tumorigenesis,and calcification process." J.Neurosci. 17. 2376-2382 (1997)
[Publications] Toshiki Tanaka: "Lipocalin-type prostaglandin D synthase (b-trace) in a newly recognized type of retinoid transporter." J.Biol.Chem.272. 15789-15795 (1997)
[Publications] Luis F.Garcia-Fernandez: "Hypothyroidism alters the expression of prostaglandin D2 synthase/β-trace in specific areas of the developing rat brain. Eur." J.Neurosci.9. 1566-1573 (1997)
[Publications] Atsushi Hiraoka: "Analysis of low-molecular-mass proteins in cerebrospinal fluid by sodium dodecyl sulfate capillary electrophoresis." J.Chromatogr.697. 141-147 (1997)
[Publications] Peter Dieter: "Arachidonic acid cascade in kupffer cells." Cells Hepatic Sinusoid. 6. 355-356 (1997)
[Publications] Yoshihide Kanaoka: "Cloning and crystal structure of hematopoietic prostaglandin D synthase." Cell. 90. 1085-1095 (1997)
[Publications] Ishtiaq Mahmud: "Prostaglandin D synthase in human megakaryoblastic cells." J.Biol.Chem.272. 28263-28266 (1997)
[Publications] Yutaka Eguchi: "Expression of lipocalin-type prostaglandin D synthase (b-trace) in human heart and its accumulation in the coronary circulation of angina patients." Proc.Natl.Acad.Sci.USA.94. 14689-14694 (1997)
[Publications] Yoshihiro Tokugawa: "Lipocalin-type prostaglandin D synthase in human male reproductive organs and seminal plasma." Biol.Reprod. 58. 600-607 (1998)
[Publications] Yoshihiro Urade: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)
[Publications] Yoshihiro Urada: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)
[Publications] Atsushi Hiraoka: "Sodium dodecyl sulfate capillary electrophoretic analysis of molecular mass heterogeneity of b-trace protein in cerebrospinal fluid from patients with central nervous system diseases." J.Chromatogr.(in press). (1998)
[Publications] Dmitry Gerashchenko: "Localization of lipocalin-type prostanoid D synthase (b-trace) in iris,cilliary body,and eye fluids." Invest.Ophthalmol.Vis.Sci.(in press). (1998)
[Publications] Robyn L.Gerena: "Identification of a fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase." Biol.Reprod.(in press). (1998)
[Publications] Luis F.Garcia-Fernandez: "Identification of a thyroid hormone response element in the 5'-flanking region of the rat lipocalin-type prostaglandin D2 synthase (b-trace) gene." Mol.Brain.Res.(in press). (1998)
[Publications] Dmitry Gerashchenko: "Dominant expression of rat prostanoid DP receptor mRNA in leptomeninges,inner segments of photoreceptor cells,iris epithelium,and cilliary processes." J.Neurochem. (in press). (1998)

1997 Fiscal Year Annual Research Report

ジーンターゲテイングによる分泌型プロスタグランジンD合成酵素の機能解析

Principal Investigator

裏出 良博 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (10201360)

Research Products

[Publications] Yoshihiro Urada: "Lipid Mediators Cell Signalling" J.Neurosci.257-273. 12. 257-273 (1995)

[Publications] Bettina Blodorn: "he major site of mRNA expression for the b-trace protein (prostaglandin D synthase) in human brain." Neurosci.Lett.209. 117-120 (1996)

[Publications] Hiroshi Oda: "Quantitative sandwich immunosorbent assay of human secretory prostaglandin D synthase (b-trace)." Proc.Japan Acad.72. 108-111 (1996)

[Publications] Carsten T.Beuckmann: "Lipocalin-type prostaglandin D synthase (b-trace) is located in pigmented epithelial cells of rat retina and accumulates within interphotoreceptor matrix." J.Neurosci.16. 6119-6124 (1996)

[Publications] Yoshihiro Urade: "Molecular mechanism of sleep regulation by prostaglandin D2. J." Lipid Mediators Cell Signalling. 14. 71-82 (1996)

[Publications] Dimitrios N.Melegos: "Immunofluorometric assay of prostaglandin D synthase (b-trace) and preliminary clinical applications." Clin.Chem.42. 1984-1991 (1996)

[Publications] Hiroyuki Toh: "IGlutathione-independent prostaglandin D synthase as a lead molecule for designing new functional proteins." Protein Engineering. 9. 1067-1082 (1996)

[Publications] Tetsumori Yamashima: "IProstaglandin D synthase (b-trace) in human arachnoid and meningioma cells : roles as a cell marker or in CSF absorption,tumorigenesis,and calcification process." J.Neurosci. 17. 2376-2382 (1997)

[Publications] Toshiki Tanaka: "Lipocalin-type prostaglandin D synthase (b-trace) in a newly recognized type of retinoid transporter." J.Biol.Chem.272. 15789-15795 (1997)

[Publications] Luis F.Garcia-Fernandez: "Hypothyroidism alters the expression of prostaglandin D2 synthase/β-trace in specific areas of the developing rat brain. Eur." J.Neurosci.9. 1566-1573 (1997)

[Publications] Atsushi Hiraoka: "Analysis of low-molecular-mass proteins in cerebrospinal fluid by sodium dodecyl sulfate capillary electrophoresis." J.Chromatogr.697. 141-147 (1997)

[Publications] Peter Dieter: "Arachidonic acid cascade in kupffer cells." Cells Hepatic Sinusoid. 6. 355-356 (1997)

[Publications] Yoshihide Kanaoka: "Cloning and crystal structure of hematopoietic prostaglandin D synthase." Cell. 90. 1085-1095 (1997)

[Publications] Ishtiaq Mahmud: "Prostaglandin D synthase in human megakaryoblastic cells." J.Biol.Chem.272. 28263-28266 (1997)

[Publications] Yutaka Eguchi: "Expression of lipocalin-type prostaglandin D synthase (b-trace) in human heart and its accumulation in the coronary circulation of angina patients." Proc.Natl.Acad.Sci.USA.94. 14689-14694 (1997)

[Publications] Yoshihiro Tokugawa: "Lipocalin-type prostaglandin D synthase in human male reproductive organs and seminal plasma." Biol.Reprod. 58. 600-607 (1998)

[Publications] Yoshihiro Urade: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)

[Publications] Yoshihiro Urada: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)

[Publications] Atsushi Hiraoka: "Sodium dodecyl sulfate capillary electrophoretic analysis of molecular mass heterogeneity of b-trace protein in cerebrospinal fluid from patients with central nervous system diseases." J.Chromatogr.(in press). (1998)

[Publications] Dmitry Gerashchenko: "Localization of lipocalin-type prostanoid D synthase (b-trace) in iris,cilliary body,and eye fluids." Invest.Ophthalmol.Vis.Sci.(in press). (1998)

[Publications] Robyn L.Gerena: "Identification of a fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase." Biol.Reprod.(in press). (1998)

[Publications] Luis F.Garcia-Fernandez: "Identification of a thyroid hormone response element in the 5'-flanking region of the rat lipocalin-type prostaglandin D2 synthase (b-trace) gene." Mol.Brain.Res.(in press). (1998)

[Publications] Dmitry Gerashchenko: "Dominant expression of rat prostanoid DP receptor mRNA in leptomeninges,inner segments of photoreceptor cells,iris epithelium,and cilliary processes." J.Neurochem. (in press). (1998)

裏出良博 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (10201360)