位置:成果数据库 > 期刊 > 期刊详情页
绿豆胰蛋白酶抑制剂精氨酸活性片段衍生物的克隆表达及生物学活性研究
  • ISSN号:0253-2395
  • 期刊名称:山西大学学报(自然科学版)
  • 时间:0
  • 页码:-
  • 分类:Q814[生物学—生物工程]
  • 作者机构:[1]山西大学生物技术研究所,化学生物学与分子工程教育部重点实验室,山西太原030006
  • 相关基金:基金项目:国家自然科学基金(31040018;31271516;8115026);国家大学生创新性实验项目基金(2012825);高等学校博士学科点专项科研基金(20111401110011)
  • 相关项目:结肠腺瘤息肉蛋白突变引发Gardner综合症及其纤毛生长紊乱之分子机制研究
中文摘要:

绿豆胰蛋白酶抑制剂(MBTI)具有赖氨酸和精氨酸两个活性中心.通过化学合成MBTI—Argc编码基因,克隆到原核表达载体pGEX-4T-1并转化到大肠杆菌DH5a.表达的可溶性融合蛋白GST—Argc经GST层析柱纯化后体外测定活性表明,该片段3.3分子对1分子胰蛋白酶活力的抑制率不足5%.DTNB法定量测定二硫键表明,与天然产物相比,Argc的二硫键个数有所减少.GST—Argc对细胞增殖和迁移生物学效应的检测实验发现,GST—Argc能显著促进细胞的增殖和迁移效应.上述结果表明,MBTI—Argc与天然产物相比在结构和功能上都发生了显著变化,表现出类生长因子活性,可能在组织细胞损伤修复及创伤愈合中具有广泛应用前景.

英文摘要:

Mung bean trypsin inhibitor contains lysine and arginine active centers. To study the biological ac- tivity of arginine active fragment (MBTI-Arg), the coding sequence of MBTI-Argc by chemical synthesis was cloned into the prokaryotic expression vector pGEX-4T-1 and transformed into E. coli DH5a. After IPTG induction,the fusion protein GST-Argc was purified by GST affinity column. The assay for trypsin inhibitory activity of GST-Argc showed that it had a weak trypsin inhibitor activity. DTNB assay was used to detect the number of free thiols. The result showed that compared with natural products,the number of disulfides were changed into two from three in Argc. Moreover,the biological effects of GST-Argc on cell proliferation and migration were detected by MTT test and scratching wound healing assay, respectively. The results indicated that GST-Argc could significantly promote cell proliferation and migration. Further- more,the proliferation effects of GST-Argc on normal cells were more significant than that of cancer cells. The findings suggest MBTI-Argc exhibits a growth factor-like activity with a relatively low trypsin inhibi- tory activity and the MBTI-Argc may play a role in cellular and tissue injury recovery.

同期刊论文项目
同项目期刊论文
期刊信息
  • 《山西大学学报:自然科学版》
  • 北大核心期刊(2011版)
  • 主管单位:山西省教育厅
  • 主办单位:山西大学
  • 主编:杨斌盛
  • 地址:太原市坞城路92号
  • 邮编:030006
  • 邮箱:xbbjb@sxu.edu.cn
  • 电话:0351-7010455
  • 国际标准刊号:ISSN:0253-2395
  • 国内统一刊号:ISSN:14-1105/N
  • 邮发代号:22-42
  • 获奖情况:
  • 边疆七年获山西省一级期刊荣誉(1993-1999)
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国动物学记录,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:5651