The investigation of the physiological function of GBP28 which is specific for adipose tissue

脂肪组织特异性GBP28的生理功能研究

• 批准号: 11672180
• 负责人: TOBE Takashi
• 金额: $ 2.24万
• 依托单位: Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672180/
• 关键词: GBP28; recombinant; amino acid; mutagenesis; セリン; 脂肪細胞特異的蛋白質; 組換え体; CHO細胞

Human GBP28, a novel plasma gelatini-binding protein, exists in a homo-trimer through its collagen-like domain and further combines to make oligomeric complexes. In order to examine the molecular basis for the complex formation, human GBP28 has been overexpressed in Chinese hamster ovary cells. The recombinant wild-type GBP28 is post-translationally processed in the same way as the native protein. Although the glycosylation of recombinant wild-type GBP28 is slightly different from native GBP28, the oligomerization of trimeric recombinant wild-type GBP28 was observed. The substitution Cys^<152> to Ser indicated that the mutated protein could not form the trimeric structural unit, and did not assemble into higher order structures. Although the mutated GBP28 DNA was transcribed as well as wild-type GBP28 DNA, the substitution decreased the amount of detectable recombinant GBP28, whose mechanism was unknown. In the case of Cys36Ser-mutated GBP28, the recombinant protein was not detected by Western-blot at all. These results strongly indicate that the Cys36 residue is essential for the polypeptide folding of GBP28 molecule..

人 GBP28 是一种新型血浆明胶结合蛋白，通过其胶原蛋白样结构域以同源三聚体形式存在，并进一步结合形成寡聚复合物。为了检查复合物形成的分子基础，人 GBP28 在中国仓鼠卵巢细胞中过表达。重组野生型 GBP28 以与天然蛋白相同的方式进行翻译后加工。尽管重组野生型GBP28的糖基化与天然GBP28略有不同，但观察到三聚体重组野生型GBP28的寡聚化。 Cys^<152>替换为Ser表明突变蛋白不能形成三聚体结构单元，并且不能组装成更高级的结构。尽管突变的 GBP28 DNA 与野生型 GBP28 DNA 一样被转录，但这种取代减少了可检测的重组 GBP28 的量，其机制尚不清楚。对于Cys36Ser突变的GBP28，Western-blot根本没有检测到重组蛋白。这些结果强烈表明 Cys36 残基对于 GBP28 分子的多肽折叠是必需的。

项目成果

Kiyomi Saito et al.: "Organization of the gene for gelatin-binding protein (GBP28)"GENE. 229. 67-73 (1999)

Kiyomi Saito 等：“明胶结合蛋白（GBP28）基因的组织”GENE。

Kiyomi Saito et al.: "Regulation of Gelatin-Binding Protein 28 (GBP28) Gene Expression by C/EBP"Biol.Pharm.Bull.. 22(11). 1158-1162 (1999)

Kiyomi Saito 等人：“C/EBP 对明胶结合蛋白 28 (GBP28) 基因表达的调节”Biol.Pharm.Bull.. 22(11)。