Interaction of arginine with protein during refolding process probed by amide H/D exchange mass spectrometry and isothermal titration calorimetry

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	Interaction of arginine with protein during refolding process probed by amide H/D exchange mass spectrometry and isothermal titration calorimetry
	Zhao, D. W. ; Liu, Y. D. ; Zhang, G. F. ; Zhang, C. ; Li, X. A. ; Wang, Q. Q. ; Shi, H. ; Su, Z. G.
刊名	Biochimica Et Biophysica Acta-Proteins and Proteomics
	2015
期号	1 页码:39-45
关键词	HDX-MS ITC Refolding Arginine rhG-CSF HYDROGEN/DEUTERIUM EXCHANGE ESCHERICHIA-COLI HYDROPHOBIC INTERACTION DISULFIDE-ISOMERASE PROPOSED MECHANISM HYDROGEN-EXCHANGE RENATURATION AGGREGATION LYSOZYME BINDING
其他题名	BBA-Proteins Proteomics
中文摘要	Arginine has been widely used as low molecular weight additive to promote protein refolding by suppressing aggregate formation. However, methods to investigate the role of arginine in protein refolding are often limited on protein's global conformational properties. Here, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was used to study the effects of arginine on recombinant human granulocyte colony-stimulating factor (rhG-CSF) refolding at the scale of peptide mapping. It was found that deuteration levels of rhG-CSF refolded with arginine was higher than that without arginine during the whole refolding process, but they became almost the same when the refolding reached equilibrium. This phenomenon indicated that arginine could protect some amide deuterium atoms from being exchanged with hydrogen, but the protection diminished gradually along with refolding proceeding. Enzymatic digestion revealed six particular peptides of 16-47,72-84, 84-93,114-124,145-153 and 154-162 were mainly responsible for the deuteration, and all of them dominantly located in protein's alpha-helix domain. Furthermore, thermodynamics analysis by isothermal titration calorimetry provided direct evidence that arginine could only react with denatured and partially refolded rhG-CSF. Taking all of the results together, we suggest that arginine suppresses protein aggregation by a reversible combination. At the initial refolding stage, arginine could combine with the denatured protein mainly through hydrogen bonding. Subsequently, arginine is gradually excluded from protein with protein's native conformation recovering. (C) 2014 Elsevier B.V. All rights reserved.
收录类别	SCI
原文出处	://WOS:000346943700005
语种	英语
WOS记录号	WOS:000346943700005
公开日期	2015-04-01
内容类型	期刊论文
源URL	[http://ir.ipe.ac.cn/handle/122111/11899]
专题	过程工程研究所_研究所（批量导入）
推荐引用方式 GB/T 7714	Zhao, D. W.,Liu, Y. D.,Zhang, G. F.,et al. Interaction of arginine with protein during refolding process probed by amide H/D exchange mass spectrometry and isothermal titration calorimetry[J]. Biochimica Et Biophysica Acta-Proteins and Proteomics,2015(1):39-45.
APA	Zhao, D. W..,Liu, Y. D..,Zhang, G. F..,Zhang, C..,Li, X. A..,...&Su, Z. G..(2015).Interaction of arginine with protein during refolding process probed by amide H/D exchange mass spectrometry and isothermal titration calorimetry.Biochimica Et Biophysica Acta-Proteins and Proteomics(1),39-45.
MLA	Zhao, D. W.,et al."Interaction of arginine with protein during refolding process probed by amide H/D exchange mass spectrometry and isothermal titration calorimetry".Biochimica Et Biophysica Acta-Proteins and Proteomics .1(2015):39-45.