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作 者:李海洁[1] 曾彬[1] 张灼寒[1] 王丽娜[1] 张竹红[1] 杨荣存[1]
机构地区:[1]南开大学医学院,天津300071
出 处:《中国生物化学与分子生物学报》2008年第5期432-438,共7页Chinese Journal of Biochemistry and Molecular Biology
基 金:国家自然科学基金(No.30540022);国家科技部基金(No.06C26211200695);国家高技术研究发展计划(863计划;No.2006AA020502);天津市科委基金(No.07JCZDJC03300;06ZHCXSH04800)~~
摘 要:SOCS3(suppressors of cytokine signaling 3)可以调节信号转导过程并影响造血干细胞的分化.利用生物信息学方法,发现SOCS3蛋白含有脯氨酸富集区,采用重组PCR技术缺失SOCS3蛋白中的129-161位的氨基酸(SOCS3Δ129-161),并将其克隆到pcDNA31/V5质粒载体中.转染细胞,然后通过流式细胞技术,分析预测的功能区对SOCS3蛋白功能的影响.结果表明,此脯氨酸富集区对SOCS3蛋白的三级结构和功能有重要的意义.建模结果显示,SOCS3Δ129-161蛋白结构发生显著变化.脯氨酸富集区的缺失也影响小鼠骨髓细胞向CD8^+T细胞分化.研究结果显示,SOCS3蛋白中129-161位的脯氨酸富集区在其结构和功能上有重要的作用.Suppressor of cytokine signaling 3 (SOCS3) negatively regulates signal transduction and exerts an important role in regulating the differentiation of hematopoietic cells. SOCS3 contains an proline-rich region as determined by bioinformatics-based analysis. To determine the role of this proline-rich region, the deletion mutant of SOCS3 amino acids 129- 161 (SOCS3△129-161) was obtainde using PCR and then cloned into pcDNA31/V5 recombinant plasmid. The functions of SOCS3△129-161were analyzed by flow cytometry in transfected cells. The results showed that the three-dimensional structure of SOCS3△129-161 was clearly different from that of wildtype as predicted by comparative(homology) computation modeling. In addition, the SOCS3△129-161 abrogated the functions of native SOCS3, inducing CD8^+ T cell differentiation. The results suggested that the proline-rich region within amino acids 129 - 161 of SOCS3 might play an important role in the formation and maintaining of SOCS3 tertiary structure and significantly contributed for its biological functions.
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