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-Fand（PEF）钙结合蛋白家族的新成员，其中包括ALG-2（凋亡链接的基因2），Calpain和Sorcin。我们准备了一种识别卵石的特定单克隆抗体。通过免疫沉淀，然后进行蛋白质印迹分析，发现卵石和ALG-2形成以钙依赖性方式解离的异二聚体。虽然卵石位于细胞质中，但ALG-2位于细胞质和核中。卵石可以通过异二聚化调节ALG-2的功能。我们在细胞粘液模具dictyostelium discoideum中鉴定了PEF蛋白，这是一种模型生物体，在其生命周期中显示了从单细胞形式到多细胞形式的转化。系统发育分析表明，ALG-2和/或卵石是保守的，并且在较高和较低的真核生物中存在普遍存在，而Calpain和Sorcin的进化却相当新。通过酵母两杂化筛选，ALG-2，卵石，ALIX和ANNEXIN XI被分离为ALG-2相互作用蛋白。 N-末端Gly/Tyr/proich Annexin的纳克斯蛋白的序列与ALIX的C末端区域相似，这表明在这些区域中存在共同结合基序

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.

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)。

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。

Y. Kitaura 他4名: "Both ALG-2 and Peflin, Penta-EF-hand(PEF)Proteins, Are Stabilized by Dimerization through Their Fifth EF-hand Regions"Archives of Biochemistry and Biophysics. (印刷中). (2002)

Y. Kitaura 和其他 4 人：“ALG-2 和 Peflin，Penta-EF-hand (PEF) 蛋白质，通过第五个 EF-hand 区域进行二聚化”，生物化学和生物物理学档案（2002 年出版）。 ）