Tetrahydrobiopterin transport through plasma membranes and regulation of nitric oxide- and monoamine-dependent signaling.

四氢生物蝶呤通过质膜转运并调节一氧化氮和单胺依赖性信号传导。

• 批准号: 10671748
• 负责人: NAKANISHI Nobuo
• 金额: $ 2.43万
• 依托单位: Meikai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671748/
• 关键词: tetrahydrobiopterin; nitric oxide synthase; dopamine; serotonin; cyclic AMP; cyclic GMP; ion transporter; vesicular monoamine transporter; nitric oxide synthase; vascular monoamine transporter; nitric oxide; membrane transport; vesicular monoa

Tetrahydrobiopterin (BH4) in the cells is an essential cofactor of nitric oxide synthase (NOS), and of tyrosine hydroxylase and tryptophan hydroxylase, rate-limiting enzymes in catecholamine and serotonin biosynthetic pathways, respectively. BH4 in the extracellular fluid is also known to induce exocytotic release of catecholamines and serotonin, and to enhance the degradation of nitric oxide (NO). Since BH4 is synthesized in the cells, BH4 transport through plasma membranes is one of the most important processes in the regulation of intra- and extracellular BH4 levels In this project, we studied the BH4 transport through plasma membrane using pheochromocytoma PC12 cells and rat basophilic leukemia RBL2H3 cells. Effect of extracellular BH4 on the cell physiology was also examined.1. BH4 outward flow (BH4 secretion) and BH4 inward flow (BH4 uptake) were performed by different transport systems, respectively. The BH4 outward flow was inhibited by reserpine and tetrabenazine, which are kn … More own to be vesicular monoamine transporters, suggesting that the transporter responsible for BH4 outward flow might be a members of a family of toxin-extruding antiporters (TEXANs).2. Outward flow of BH4 from PC12 cells was down-regulated by both cyclic AMP and cyclic GMP, and protein phosphorylation was suggested to be involved in this down-regulation.3. Although PC12 and RBL2H3 cells can take up BH4 in the medium, BH4 thus incorporated was strictly distinguished from the endogenous BH4 by the cells : BH4 of the exogenous origin was quickly oxidized and subsequently excreted to the medium. Therefore, the addition of sepia pterin, an intermediate in the BH4 salvage pathway, into the medium raised cellular BH4 level more efficiently than BH4 addition into the medium.4. Effects of extracellular BH4 on the phosphorylation of ion transporters are now being examined, since we found that Na+,K+-ATPase and Na+-K+-2CI- cotransporter were regulated by the phosphorylation of the protein molecules and since BH4 was expected to induce intracellular Ca2+ elevation which can trigger the protein phosphorylation reactions. Less

细胞中的四氢生物蝶呤（BH4）是一氧化氮合酶（NOS）、酪氨酸羟化酶和色氨酸羟化酶的重要辅助因子，分别是儿茶酚胺和血清素生物合成途径中的限速酶。已知细胞外液中的BH4还可诱导儿茶酚胺和血清素的胞外释放，并增强一氧化氮（NO）的降解。由于BH4是在细胞内合成的，因此BH4在质膜上的转运是调控细胞内外BH4水平的重要过程之一。本项目以嗜铬细胞瘤PC12细胞和大鼠嗜碱性白血病RBL2H3细胞为研究对象，研究了BH4在质膜上的转运。细胞外BH4对细胞生理的影响也进行了研究。BH4向外流动（BH4分泌）和向内流动（BH4摄取）分别通过不同的转运系统进行。利血平和丁苯那嗪可抑制BH4向外流动，这两种转运蛋白通常被认为是水泡单胺转运蛋白，提示BH4向外流动的转运蛋白可能是毒素挤出反转运蛋白（TEXANs）家族的一员。环AMP和环GMP均可下调PC12细胞的BH4向外流动，且可能与蛋白磷酸化有关。虽然PC12和RBL2H3细胞可以在培养基中吸收BH4，但细胞严格区分这种吸收的BH4与内源性的BH4：外源性的BH4被迅速氧化并随后排泄到培养基中。因此，在培养基中添加sepia pterin（一种BH4修复途径的中间体）比在培养基中添加BH4更有效地提高了细胞BH4水平。目前正在研究细胞外BH4对离子转运蛋白磷酸化的影响，因为我们发现Na+，K+- atp酶和Na+-K+- 2ci -共转运蛋白受到蛋白质分子磷酸化的调节，并且BH4有望诱导细胞内Ca2+升高，从而触发蛋白质磷酸化反应。少

项目成果

Hattori,Y.: "Tetrahydrobiopterin synthesis in rat cardiac myocytes; an event required for cytokine-induced NO generation" Pteridines. 9(1). 8-12 (1998)

Hattori,Y.：“大鼠心肌细胞中四氢生物蝶呤的合成；细胞因子诱导的 NO 生成所需的事件”蝶啶。

Nakazawa, N.: "6R-erythro. tetrahydrobiopterin triggers serotonin release with 5HT-loaded RBL2H3 cells, a rat mast cell line"Pteridines. 10. 1-4 (1999)

Nakazawa, N.：“6R-赤型四氢生物蝶呤触发负载 5HT 的 RBL2H3 细胞（一种大鼠肥大细胞系）释放血清素”蝶啶。

Kurihara K. et al.: "Regulation of Na+-K+-ATPase by cAMP-dependent protein kinase anchored on membrane via its anchoring protein"Am. J. Physiol.. 279. C1516-C1527 (2000)

Kurihara K. 等人：“通过锚定蛋白锚定在膜上的 cAMP 依赖性蛋白激酶对 Na -K -ATP 酶的调节”Am。

Hasegawa, H. et al.: "Ability of RBL2H3 cells to lower environmental tetrahydrobiopterin concentration"Pteridines.. 11. 121-125 (2000)

Hasekawa, H. 等人：“RBL2H3 细胞降低环境四氢生物蝶呤浓度的能力”Pteridines.. 11. 121-125 (2000)

Kurihara, K.: "Thyroid hormone (3,5,3'-triiodo-L-thyronine) masking/inversion of stimulatory effect of androgen on expression of mk1,a true kallikrein, in the mouse submandibular gland"Endocrinology. 140. 3003-3011 (1999)

Kurihara, K.：“甲状腺激素（3,5,3-三碘-L-甲状腺氨酸）掩盖/逆转雄激素对小鼠颌下腺中 mk1（一种真正的激肽释放酶）表达的刺激作用”内分泌学。