Nerve growth factor-induced increase in biopterin level of pheochromocytoma PC12.

神经生长因子诱导嗜铬细胞瘤 PC12 生物蝶呤水平增加。

• 批准号: 62570841
• 负责人: NAKANISHI Nobuo
• 金额: $ 1.15万
• 依托单位: Meikai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570841/
• 关键词: Nerve growth factor; Pheochromocytoma PC12; Tetrahydrobiopterin; Protein synthesis; RNA synthesis; GTP cyclohydrolase; Protein phosphorylation; Neuronal differentiation; 褐色細胞腫細胞PCl2; 神経細胞の分化; プロテインキナーゼ; K252a; 細胞内Caイオン; 細胞外ATP; 褐色細胞腫PC12; ビ

Nerve growth factor (NGF) is a protein required for the survival and development of various neurons. In culture system, NGF induces differentiation of pheochromocytoma PC12 into sympathetic neuron-like cells from chromaffin cell-like phenotype. In this study, tetrahydrobiopterin (BPH4) in PC12 cells are found to be transiently increased by NGF. Then the mechanism of this NGF action and the biological meaning of the phenomenon are investigated.1. Mechanism of action: (1) NGF-induced increase in BPH4 was blocked by actinomycin D (AD) and cycloheximide (CH). (2) The activity of GTP cyclohydrolase (GTPCH), which is thought to be a regulatory enzyme in BPH4 biosynthesis, was also transiently increased by NGF. The GTPCH activity and BPH4 level showed quite similar responses to NGF in the time course and dose dependence experiments. The NGF effect on GTPCH was also blocked by AD and 'CH. These results suggest that NGF increase BPH4 level by increasing the amount of GTPCH via synthesis of RNA and of protein. (3) Protein kinase inhibitors, H-7 and H-8, of the concentrations around K_i values for protein kinase A, protein kinase G and protein kinase C did not affect the NGF action. (4) Dibutyryl cyclic AMP (dBcAMP) also increased the GTPCH activity and BPH4 level, but the action mechanism of NGF seems to be different from that of dBcAMP.2. Biological meaning: Effect of an analog of BPH4, tetrahydro-6-methylpterin (MPH4), on the phosphorylation of a nuclear protein named SMP, of which phosphorylation is stimulated by NGF, was examined. MPH4 inhibited the NGF-stimulated phosphorylation of SMP, suggesting a possible function of BPH4 as a modulator of NGF action. Further study, however, should be needed to clarify the biological role of transient increase in BPH4 induced by NGF.

神经生长因子(NGF)是多种神经元生存和发育所必需的蛋白质。在培养体系中，NGF诱导嗜铬细胞瘤PC12细胞由嗜铬细胞表型向交感神经元样细胞分化。在这项研究中，发现四氢生物蝶呤(BPH4)在PC12细胞中被发现被NGF瞬时增加。并对NGF作用的机制和该现象的生物学意义进行了探讨。作用机制：(1)放线菌素D(AD)和放线菌亚胺(CH)可阻断NGF诱导的BPH4升高。(2)被认为是BPH4生物合成调节酶的GTP环水解酶(GTPCH)的活性也被NGF瞬时上调。在时间进程和剂量依赖实验中，GTPCH活性和BPH4水平对NGF的反应非常相似。AD和‘CH也可阻断NGF对GTPCH的作用。这些结果表明，NGF通过合成RNA和蛋白质而增加GTPCH的含量，从而提高BPH4的水平。(3)K_I值附近的蛋白激酶抑制剂H-7和H-8对NGF的作用无明显影响。(4)二丁基环AMP(DBcAMP)也能提高GTPCH活性和BPH4水平，但NGF的作用机制似乎与dBcAMP不同。生物学意义：研究了BPH4的类似物四氢-6-甲基蝶呤(MPH4)对一种名为SMP的核蛋白的磷酸化的影响，该蛋白的磷酸化是由NGF刺激的。MPH4抑制NGF刺激的SMP的磷酸化，提示BPH4可能是NGF作用的调节器。然而，需要进一步的研究来阐明NGF诱导的BPH4一过性增加的生物学作用。

项目成果

中川幹也: "Teratoma 402AX細胞のNADH特異性ジヒドロプテリジン還元酵素に対するretinoic acid,dibutyryl cyclic AMP、および神経成長因子の影響" 明海大学歯学雑誌.

Mikiya Nakakawa：“视黄酸、二丁酰环AMP和神经生长因子对畸胎瘤402AX细胞中NADH特异性二氢蝶啶还原酶的影响”明海大学牙科杂志。

Nobuo NAKANISHI: ""Enzymes of Glycolytic Pathway.Pyruvate kinase,phosphofructokinase,and lactate dehydrogenase"in Dynamic Aspect of Dental Pulp" Ellis Horwood.,

Nobuo NAKANISHI：“牙髓动态方面的糖酵解途径酶。丙酮酸激酶、磷酸果糖激酶和乳酸脱氢酶”Ellis Horwood。，

Nobuo NAKANISHI: "Dynamic Aspects of Dental Pulp." Ellis Horwood, England,

Nobuo NAKANISHI：“牙髓的动态方面。”

大谷徹: "神経成長因子によるpheochromocytoma PC12h細胞の神経突起形成誘導作用に対するアルコ-ルの影響" 明海大学歯学雑誌.

Toru Otani：“酒精对神经生长因子诱导嗜铬细胞瘤 PC12h 细胞神经突形成的影响”明海大学牙科杂志。

Hiroyuki HASEGAWA: "“Dihydropteridine reductase" in Methods in Enzymology Vo.142 (Kaufman,S.,ed.) pp.103ー110." Academic Press,New York, (1987)

Hiroyuki HASEGAWA：“酶学方法第 142 卷中的“二氢蝶啶还原酶”（Kaufman，S. 编辑）第 103-110 页，纽约学术出版社，（1987 年）