Molecular Mechanism of production and secretion of platelet-derived bioactive lysolipids

血小板源性生物活性溶血脂产生和分泌的分子机制

• 批准号: 13480198
• 负责人: IGARASHI Yasuyuki
• 金额: $ 9.6万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13480198/
• 关键词: bioactive lipids; sphingosine1-phosphate; Edg Receptors; lysophosphatidic acid; ABC transporter; sphingosine kinase; alycolipids; lipid signaling; リゾボスファチジン酸; GPCR

The receptors for serum-borne lysophospholipid mediators such as sphingosine1-phosphate (S1P) and lysophosphatidic acid (LPA) was identified in these several years and their physiological and pathophysiological roles have been strongly concerned. These bioactive lipids were observed to be released from activated platelets, but the detailed mechanism of their production, secretion and degradation have not been thoroughly studied. Therefore, in this three years Japan-USA research project, we have tried to clarify the molecular mechanism of production, secretion, and degradation of these bioactive lysolipids in the strong collaboration between Japanese and American groups.Through the collaboration we tried and found the following results. (1) Blood S1P is derived mainly from stimulated platelets and LPA is partially released from stimulated platelets but mostly produced in the blood by phospholipases which are released from activated platelets with the aids of lysophospholipase D. (2) We partially purified the responsible phospholipases which produce the lysophospholipids from the blood phospholipids. (3) The involvement of ABC transporter in S1P releasing from platelets was found but we could not determine the specific ABC transporter involved. (4) We identified for the first time a sphingolipid transporter RSB 1, a ATP-dependent new type of six transmembrane transporter, which can exclude the over-loaded DHS and PHS from yeast cells (5) We determined the molecular mechanism of ligand-receptor interaction utilizing computational model of Edg-6 (S1P6)/S1P interaction.By these collaboration studies we enriched our knowledge how the metabolism and functions of the blood-borne bioactive lysolipids such as LPA and S1P are regulated, indicating the importance of the regulations of these lysolipids in blood vessel biology and pathology.

近年来，血清溶血磷脂介质如 1-磷酸鞘氨醇 (S1P) 和溶血磷脂酸 (LPA) 的受体被鉴定出来，它们的生理和病理生理作用受到了强烈关注。观察到这些生物活性脂质从活化的血小板中释放出来，但其产生、分泌和降解的详细机制尚未得到彻底研究。因此，在这个为期三年的日美研究项目中，我们试图在日本和美国研究小组的大力合作下阐明这些生物活性溶血脂的产生、分泌和降解的分子机制。通过合作，我们尝试并发现了以下结果。 (1) 血液中的 S1P 主要来源于受刺激的血小板，LPA 部分从受刺激的血小板中释放出来，但大部分是由磷脂酶在血液中产生的，磷脂酶是在溶血磷脂酶 D 的帮助下从活化的血小板中释放出来的。 (2) 我们部分纯化了负责从血液磷脂中产生溶血磷脂的磷脂酶。 (3)发现ABC转运蛋白参与血小板释放S1P，但无法确定具体参与的ABC转运蛋白。 （4）我们首次鉴定了鞘脂转运蛋白RSB 1，一种ATP依赖性新型六跨膜转运蛋白，可以排除酵母细胞中超载的DHS和PHS。（5）我们利用Edg-6（S1P6）/S1P相互作用的计算模型确定了配体-受体相互作用的分子机制。通过这些合作研究，我们丰富了我们的知识 LPA、S1P等血源性生物活性溶血脂的代谢和功能是如何调控的，表明这些溶血脂的调控在血管生物学和病理学中的重要性。

项目成果

Kihara A.: "Identification and characterization of a saccharomyces cerevisiae gene, RSB1, involved in sphingoid long-chain base release."J.Biol.Chem.. 277. 30048-30054 (2002)

Kihara A.：“参与鞘氨醇长链碱基释放的酿酒酵母基因 RSB1 的鉴定和表征。”J.Biol.Chem.. 277. 30048-30054 (2002)

Kihara A., Igarashi Y.: "Identification and characterization of a saccharomyces cerevisiae gene, RSB1, involved in sphingoid long-chain base release."J.Biol.Chem.. 277. 30048-30054 (2002)

Kihara A.、Igarashi Y.：“参与鞘氨醇长链碱基释放的酿酒酵母基因 RSB1 的鉴定和表征。”J.Biol.Chem.. 277. 30048-30054 (2002)

Takashi Murate: "Cell Type-specific Localization of Sphingosine Kinase la in Human Tissues"The Journal of Histochemistry & Cytochemistry. 49(7). 845-855 (2001)

Takashi Murate：“人体组织中鞘氨醇激酶 la 的细胞类型特异性定位”组织化学杂志

Sano T.: "Novel mechanism for lysophosphatidic acid production involving activated platelets"J. Biol. Chem.. 227・24. 21197-21206 (2002)

Sano T.：“涉及活化血小板的溶血磷脂酸产生的新机制”J. Biol. 227・24（2002）。

Ogawa C.: "Identification and characterization of a novel human sphingosine 1-phosphate phosphohydrolase, hSPP2"J. Biol. Chem.. 278. 1268-1272 (2003)

小川 C.：“新型人鞘氨醇 1-磷酸磷酸水解酶 hSPP2 的鉴定和表征”J。