Over-expression and refolding of isotopically labeled recombinant catalytic domain of human macrophage elastase (MMP-12) for NMR studies

CORC > 上海有机化学研究所 > 中国科学院上海有机化学研究所 > 上海有机化学研究所

	Over-expression and refolding of isotopically labeled recombinant catalytic domain of human macrophage elastase (MMP-12) for NMR studies
	Zheng XH(郑训海) ; Ou L(欧力) ; Tong XT(童孝田) ; Zhu J(朱静) ; Wu HM(吴厚铭)
刊名	Protein Expr. Purif.
	2007
卷号	56 期号:2 页码:160-166
ISSN号	1046-5928
其他题名	用于NMR 研究的基质金属蛋白酶MMP12催化域重组子的表达和重折叠
通讯作者	吴厚铭
英文摘要	Human macrophage elastase (MMP-12) plays an important role in inflammatory processes and is involved in a number of physiological or pathological situations, such as conversion of plasminogen into angiotatin, allergic airway inflammation, vascular remodeling or alteration, as well as emphysema, and has been justified as a novel drug target. Here, we report the over-expression in Escherichia coil, purification and refolding of MMP-12 catalytic domain for NMR studies. The primary sequence of expressed protein was identified by means of MALDI-TOF MS, and was confirmed by the MALDI-TOF MS data of trypsin-digested peptides. A significantly optimized protocol has been worked out to prepare N-15 and/or C-13-labeled MMP-12 catalytic domain, and the yield of the purified protein is estimated to 10-12 mg from 0.5 L of M9 minimal media. Finally, the N-15-H-1 HSQC spectrum of uniformly N-15-labeled MMP-12 catalytic domain indicates the presence of well-ordered and properly folded protein in a monomeric form. (C) 2007 Elsevier Inc. All rights reserved.
学科主题	生命有机化学
收录类别	SCI
原文出处	http://dx.doi.org/10.1016/j.pep.2007.05.005
语种	英语
WOS记录号	WOS:000251312300003
公开日期	2013-02-21
内容类型	期刊论文
源URL	[http://202.127.28.38/handle/331003/17383]
专题	上海有机化学研究所_上海有机化学研究所
推荐引用方式 GB/T 7714	Zheng XH,Ou L,Tong XT,et al. Over-expression and refolding of isotopically labeled recombinant catalytic domain of human macrophage elastase (MMP-12) for NMR studies[J]. Protein Expr. Purif.,2007,56(2):160-166.
APA	郑训海,欧力,童孝田,朱静,&吴厚铭.(2007).Over-expression and refolding of isotopically labeled recombinant catalytic domain of human macrophage elastase (MMP-12) for NMR studies.Protein Expr. Purif.,56(2),160-166.
MLA	郑训海,et al."Over-expression and refolding of isotopically labeled recombinant catalytic domain of human macrophage elastase (MMP-12) for NMR studies".Protein Expr. Purif. 56.2(2007):160-166.