Studies on ghe novel neurotrophic factor, secretory phospholiase A_2

新型神经营养因子分泌型磷酸酶A_2的研究

• 批准号: 14560058
• 负责人: ARIOKA Manabu
• 金额: $ 2.3万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560058/
• 关键词: secretory phospholiase A_2; neurotrophic factor; L-type Ca^<2+> channel; apoptosis; PC12; cerebellar granule neuron; neurite outgrowth

Secretory phospholipases A_2(sPLA_<2s> belong to a broad and structurally diverse family of enzymes that hydrolyse the sn-2 ester bond of glycerophospholipids. We previously showed that a secreted fungal 15kDa protein, named p15, as well as its ortholog from Streptomyces coelicolor (named Scp15) induce neurite outgrowth in PC12 cells at nanomolar concentrations. We report here that both p15 and Scp15 are members of a newly identified group of fungal/bacterial sPLA_<2s>. The phospholipids hydrolyzing activity of p15 is absolutely required for neurite outgrowth induction. Mutants with a reduced PLA_2 activity exhibited a comparable reduction in neurite-inducing activity, and the ability to induce neuritis closely matched the capacity of various p15 forms to promote fatty acid release from livePC12 cells. A structurally divergent member of the sPLA_2 family, bee venom sPLA_2, also induced neuritis in a phospholipase activity-dependent manner, and the same effect was elicited by mouse group V and X sPLA_<2s>, but not by group IB and IIAsPLA_<2s>. Lysophosphatidylcholine, but not other lysophospholipids, nor arachidonic acid, elicited neurite outgrowth in an L-type Ca^<2+> channel activity-dependent manner. In addition, p15-induced neuritogenesis was unaffected by various inhibitors that block arachidonic acid conversion into bioactive eicosanoids. Altogether, these results delineate a novel, Ca^<2+>-and lysophosphatidylcholine-dependent neurotrophin-like role of sPLA_<2s> in the nervous system.

分泌型磷脂酶A_2(Spla_&lt；2s&gt；)是一个广泛的、结构多样的酶家族，它能分解甘油磷脂的sn-2酯键。我们之前的研究表明，天蓝色链霉菌分泌的15 kDa蛋白p15及其同源物Scp15在纳摩尔浓度下诱导PC12细胞突起生长。我们在此报告，p15和Scp15都是一个新发现的真菌/细菌Spla_&lt；2s&gt；组的成员。P15的磷脂水解性是诱导神经突起生长所必需的。PLA2活性降低的突变体显示出类似的轴突诱导活性降低，并且诱导神经炎的能力与各种p15形式促进活体PC12细胞释放脂肪酸的能力非常接近。SPLA2家族中结构不同的成员蜂毒SPLA2也以一种磷脂酶活性依赖的方式诱导神经炎，V组和X SPLA2组小鼠也有同样的作用，而IB组和IIASPLA2组则不能。溶血磷脂酰胆碱，而不是其他溶血磷脂，也不是花生四烯酸，以L型钙通道活性依赖的方式引起轴突生长。此外，p15诱导的神经再生不受阻止花生四烯酸转化为生物活性二十烷酸类化合物的各种抑制剂的影响。综上所述，这些结果描述了SPLA2S和GT；在神经系统中的一个新的钙离子和溶血磷脂酰胆碱依赖的神经营养素样作用。

项目成果

Nakashima, S., et al.: "Identification and characterization of Scp15, a novel protein from Streptomyces coelicolor A3(2) with neurite-inducing activity in PC12 cells."Biosci.Biotech.Biochem.. 67(1). 77-82 (2003)

Nakashima, S. 等人：“Scp15 的鉴定和表征，Scp15 是一种来自天蓝色链霉菌 A3(2) 的新型蛋白质，在 PC12 细胞中具有神经突诱导活性。”Biosci.Biotech.Biochem.. 67(1)。

Nakashima, S., et al.: "Secretory phospholipases A_2 induce neurite outgrowth in PC12 cells."Biochem.J.. 376(3). 655-666 (2003)

Nakashima, S., et al.：“分泌性磷脂酶 A_2 诱导 PC12 细胞中的神经突生长。”Biochem.J. 376(3)。

Oka, M., et al.: "Molecular cloning and functional characterization of avaB, a gene encoding Vam6p/Vps39p-like protein in Aspergillus nidulans."FEMS Microbiol.Lett.. 232・1. 113-121 (2004)

Oka, M., et al.：“avaB 的分子克隆和功能表征，avaB 是曲霉中编码 Vam6p/Vps39p 样蛋白的基因。”FEMS Microbiol.Lett.. 232・1 (2004)。

Nakashima, S., et al.: "Functional expression in Aspergillus oryzae of p15, a protein with potent neurite-inducing activity in PC12 cells."Biosci.Biotech.Biochem.. 66・3. 674-678 (2002)

Nakashima, S., et al.：“p15 在米曲霉中的功能表达，p15 是一种在 PC12 细胞中具有有效神经突诱导活性的蛋白质。”Biosci.Biotech.Biochem.. 66・3 (2002)。

Ohsumi, K., et al.: "Cloning and characterization of a gene (avaA) from Aspergillus nidulans encoding a small GTPase involved in vacuolar biogenesis."Gene. 291(1-2). 77-84 (2002)

Ohsumi, K. 等人：“来自构巢曲霉的基因 (avaA) 的克隆和表征，该基因编码参与液泡生物发生的小 GTP 酶。”基因。