Functional analysis of a novel intracellular calcium-binding protein peflin

新型细胞内钙结合蛋白peflin的功能分析

• 批准号: 11460038
• 负责人: MAKI Masatoshi
• 金额: $ 7.94万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11460038/
• 关键词: peflin; ALG-2; calcium; EF-hand; interaction; Alix; annexin; dimer; カルシウム結合タンパク質; penta-EF-hand; peflin; ヘテロダイマー; アポトーシス; cDNA; EF-hand

Peflin is a new member of the penta-EF-hand (PEF) calcium-binding protein family that includes ALG-2 (apoptosis linked gene 2), calpain, and sorcin. We prepared a specific monoclonal antibody that recognized peflin. By immunoprecipitation followed by Western blot analysis, peflin and ALG-2 have been found to form a heterodimer that dissociates in a calcium-dependent manner. While peflin is localized to cytoplasm, ALG-2 is localized in both cytoplasm and nuclei. Peflin may regulate function of ALG-2 by heterodimerization. We identified PEF proteins in the cellular slime mold Dictyostelium discoideum, a model organism that displays transformation from unicellular form to multicellular form during its life cycle. Phylogenetic analysis has revealed that ALG-2 and/or peflin are conserved and present ubiquitously in higher and lower eukaryotes while calpain and sorcin are rather new in evolution. By the yeast two-hybrid screening, ALG-2, peflin, Alix and annexin XI were isolated as ALG-2-interacting proteins. The N-terminal Gly/Tyr/Pro-rich region of annexin has a sequence similar to that of the C-terminal region of Alix, suggesting presence of a common binding motif in these regions

Peflin 是 penta-EF-hand (PEF) 钙结合蛋白家族的新成员，该家族包括 ALG-2（凋亡相关基因 2）、钙蛋白酶和 sorcin。我们制备了识别 peflin 的特异性单克隆抗体。通过免疫沉淀和蛋白质印迹分析，发现 peflin 和 ALG-2 形成异二聚体，以钙依赖性方式解离。 peflin 定位于细胞质，而 ALG-2 定位于细胞质和细胞核。 Peflin 可能通过异二聚化调节 ALG-2 的功能。我们在细胞粘菌盘基网柄菌中鉴定了 PEF 蛋白，盘基网柄菌是一种模式生物，在其生命周期中表现出从单细胞形式向多细胞形式的转变。系统发育分析表明，ALG-2 和/或 peflin 是保守的，普遍存在于高等和低等真核生物中，而 calpain 和 sorcin 在进化中相当新。通过酵母二杂交筛选，分离出ALG-2、peflin、Alix和annexin XI作为ALG-2相互作用蛋白。膜联蛋白的 N 端富含 Gly/Tyr/Pro 的区域具有与 Alix 的 C 端区域相似的序列，表明这些区域中存在共同的结合基序

项目成果

Ohkouchi, S. et al. (6 authors): "Identification and characterization of two penta-EF-hand Ca^<2+>-binding proteins in Dictyostelium discoideum"Journal of Biochemistry. 130. 207-215 (2001)

Ohkouchi，S.等人。

Kitaura,Y. et al.: "Peflin, a novel member of the five-EF-hand-protein family, is similar to the apoptosis-linked gene 2(ALG-2) protein but possesses nonapeptide repeats in the N-terminal hydrophobic region"Biochem. Biophys. Res. Commun.. 263. 68-75 (1999

北浦，Y.

Satoh, H. et al. (5authors): "ALG-2 Interacts with the amino-terminal domain of annexin XI in a Ca^<2+>- dependent manner"Biochemical and Biophysical Research Communications. 291. 1166-1172 (2002)

佐藤，H.等人。

J.Takano 他3名: "Four Types of Calpastatin Isoforms with Distinct Amino-Terminal Sequences Are Specified by Alternative First Exons and Differentially Expressed in Mouse Tissues"Journal of Biochemistry. 128. 83-92 (2000)

J. Takano 和其他 3 人：“具有不同氨基末端序列的四种类型的钙蛋白酶抑制剂亚型由替代第一外显子指定并在小鼠组织中差异表达”《生物化学杂志》128. 83-92 (2000)。

Y.Kitaura 他5名: "Peflin, a novel member of the five-EF-hand-protein family, is similar to the apoptosis-linked gene-2 (ALG-2) protein but possesses nonapeptide repeats in the N-terminal hydrophobic region"Biochemical and Biophysical Research Communication.

Y.Kitaura 和其他 5 人：“Peflin 是五手蛋白家族的新成员，与凋亡相关基因 2 (ALG-2) 蛋白相似，但在 N- 中具有九肽重复序列。末端疏水区”生物化学和生物物理研究通讯。