Studies on the structure and function of protein kinase PKN

蛋白激酶PKN的结构与功能研究

• 批准号: 08458230
• 负责人: ONO Yoshitaka
• 金额: $ 1.34万
• 依托单位: KOBE UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08458230/
• 关键词: protein kinases; PKN; Signal transduction

To understand the physiological function of PKN,PKN-associated proteins were analyzed.1) Identification and characterization of PKN-associated proteinsPKN-assocaited proteins were identified by the yeast two-hybrid system. By this screening, it has been found that PKN asscoiates some cytoskeletal components such as neurofilament, vimentin, and a-actinin. Moreover, the interaction between PKN and PCD17 has been shown. PCD17 is a neural antigen, which is recognized by anti-Pukinje cell autoantibodies, detected in sera and cerebrospinal fluids of patients with paraneoplastic cerebellar degeneration syndrome, and it has been suggested to be transcriptional factor by the primary structure.2) Translocation of PKN from the cytosol to the nucleus induced by stressesEffects of environmental stresses on the subcellular location of PKN were investigated in the cultured cells. The immunoflueorescence of PKN resided prominently in the cytoplasmic region in non-stressed cells. When these cells were treated at the high degree of temperature, there was a time-dependent decrease of the immunofluorescence of PKN in the cytoplasmic region that correlated with an increase within the nucleus.These results described as above raise the possibilities that PKN has important roles both in the cytoskeletal organization and nuclear events.

为了解PKN的生理功能，对PKN相关蛋白进行了分析。1）PKN相关蛋白的鉴定与鉴定利用酵母双杂交系统对PKN相关蛋白进行了鉴定。通过这种筛选，已经发现PKN与一些细胞骨架成分如神经丝、波形蛋白和α-辅肌动蛋白相关。此外，PKN和PCD 17之间的相互作用已被证明。PCD 17是一种神经元抗原，在副肿瘤性小脑变性综合征患者血清和脑脊液中被抗Pukinje细胞自身抗体识别，根据其一级结构推测其可能是一种转录因子。2）应激诱导PKN从胞浆向核转位研究了环境应激对PKN亚细胞定位的影响。在非应激细胞中，PKN的免疫荧光主要位于胞浆区。当这些细胞在高温下处理时，PKN的免疫荧光在细胞质区域随时间的推移而减少，而在细胞核内则增加，这些结果表明PKN在细胞骨架的组织和核事件中起重要作用。

项目成果

Konishi, H.: "Activation of protein kinase B(Akt/RAC-protein kinase)by cellular stress and its association with heat shock protein Hsp27." FEBS Lett.410. 493-498 (1997)

Konishi, H.：“细胞应激激活蛋白激酶 B（Akt/RAC-蛋白激酶）及其与热休克蛋白 Hsp27 的关联。”

Takanaga, H.: "PKN interacts with a paraneoplastic cerebellar degeneration-associated antigen,which is a potential transcription factor." Exp.Cell Res.(in press).

Takanaga, H.：“PKN 与副肿瘤性小脑变性相关抗原相互作用，这是一种潜在的转录因子。”

Kitagawa, M.: "The role of the unique motifs in the amino-terminal region of PKN on its enzymatic" Biochem.Biophys.Res.Commun.220. 963-968 (1996)

Kitakawa, M.：“PKN 氨基末端区域的独特基序对其酶促作用的作用”Biochem.Biophys.Res.Commun.220。

Watanabe, G.: "Protein kinase N(PKN) and PKN-related protein rhophilin as targets of small GTPase rho." Science. 271. 645-648 (1996)

Watanabe, G.：“蛋白激酶 N (PKN) 和 PKN 相关蛋白 rhophilin 作为小 GTPase rho 的靶标。”

Matsuzawa,K.: "Domain-specific phosphorylation of vimentin and glial fibrillary acidic protein by PKN." Biochem.Biophys.Res.Commun.234. 621-625 (1997)

Matsuzawa,K.：“PKN 对波形蛋白和胶质纤维酸性蛋白进行域特异性磷酸化。”