An intramolecular disulfide bond designed in myoglobin fine-tunes both protein structure and peroxidase activity

doi:10.1016/j.abb.2016.04.012

	An intramolecular disulfide bond designed in myoglobin fine-tunes both protein structure and peroxidase activity
	Wu, Lei-Bin 2; Yuan, Hong 1; Zhou, Hu3 ; Gao, Shu-Qin 4; Nie, Chang-Ming 2; Tan, Xiangshi 1; Wen, Ge-Bo 4; Lin, Ying-Wu 2,4
刊名	ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
	2016-06-15
卷号	600 页码:47-55
关键词	Heme proteins Protein design Disulfide bond Coordination state Peroxidase activity
ISSN号	0003-9861
DOI	10.1016/j.abb.2016.04.012
文献子类	Article
英文摘要	Disulfide bond plays crucial roles in stabilization of protein structure and in fine-tuning protein functions. To explore an approach for rational heme protein design, we herein rationally introduced a pair of cysteines (F46C/M55C) into the scaffold of myoglobin (Mb), mimicking those in native neuroglobin. Molecular modeling suggested that it is possible for Cys46 and Cys55 to form an intramolecular disulfide bond, which was confirmed experimentally by ESI-MS analysis, DTNB reaction and CD spectrum. Moreover, it was shown that the spontaneously formed disulfide bond of Cys46-Cys55 fine-tunes not only the heme active site structure, but also the protein functions. The substitution of Phe46 with Ser46 in F46S Mb destabilizes the protein while facilitates H2O2 activation. Remarkably, the formation of an intramolecular disulfide bond of Cys46-Cys55 in F46C/M55C Mb improves the protein stability and regulates the heme site to be more favorable for substrate binding, resulting in enhanced peroxidase activity. This study provides valuable information of structure-function relationship for heme proteins regulated by an intramolecular disulfide bond, and also suggests that construction of such a covalent bond is useful for design of functional heme proteins. (C) 2016 Elsevier Inc. All rights reserved.
资助项目	National Science Foundation of China[31370812] ; National Science Foundation of China[21472027] ; National Science Foundation of China[21375138] ; National Science Foundation of China[11275090] ; Hunan Provincial Natural Science Foundation for Distinguished Young Scholars[2015JJ1012] ; Zhengxiang scholar program of the University of South China[2014-003]
WOS关键词	SPERM-WHALE-MYOGLOBIN ; RECOMBINANT MANGANESE PEROXIDASE ; DISTAL HISTIDINE ; CIRCULAR-DICHROISM ; MOLECULAR-DYNAMICS ; ARTIFICIAL ENZYMES ; CRYSTAL-STRUCTURE ; LIGAND-BINDING ; HEME ; NEUROGLOBIN
WOS研究方向	Biochemistry & Molecular Biology ; Biophysics
语种	英语
出版者	ELSEVIER SCIENCE INC
WOS记录号	WOS:000376812600005
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/276002]
专题	分析化学研究室
通讯作者	Lin, Ying-Wu
作者单位	1.Fudan Univ, Inst Biomed Sci, Shanghai Key Lab Chem Biol Prot Res, Dept Chem, Shanghai 200433, Peoples R China; 2.Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Peoples R China; 3.Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, Dept Analyt Chem, Shanghai 201203, Peoples R China; 4.Univ South China, Lab Prot Struct & Funct, Hengyang 421001, Peoples R China
推荐引用方式 GB/T 7714	Wu, Lei-Bin,Yuan, Hong,Zhou, Hu,et al. An intramolecular disulfide bond designed in myoglobin fine-tunes both protein structure and peroxidase activity[J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS,2016,600:47-55.
APA	Wu, Lei-Bin.,Yuan, Hong.,Zhou, Hu.,Gao, Shu-Qin.,Nie, Chang-Ming.,...&Lin, Ying-Wu.(2016).An intramolecular disulfide bond designed in myoglobin fine-tunes both protein structure and peroxidase activity.ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS,600,47-55.
MLA	Wu, Lei-Bin,et al."An intramolecular disulfide bond designed in myoglobin fine-tunes both protein structure and peroxidase activity".ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 600(2016):47-55.