Crystal structures of multicopper oxidase CueO G304K mutant: structural basis of the increased laccase activity

doi:10.1038/s41598-018-32446-7

	Crystal structures of multicopper oxidase CueO G304K mutant: structural basis of the increased laccase activity
	Wang HQ(王汉潜); Gong Y(宫宇); Dong YH(董宇辉); Gao ZQ(高增强); Yan YH(闫玉华); Gong, Y; Wu, NF; Tian, J; Fan, YL; Zhang, ZY
刊名	SCIENTIFIC REPORTS
	2018
卷号	8 页码:14252
ISSN号	2045-2322
DOI	10.1038/s41598-018-32446-7
文献子类	Article
英文摘要	The multicopper oxidase CueO is involved in copper homeostasis and copper (Cu) tolerance in Escherichia coli. The laccase activity of CueO G304K mutant is higher than wild-type CueO. To explain this increase in activity, we solved the crystal structure of G304K mutant at 1.49 angstrom. Compared with wild-type CueO, the G304K mutant showed dramatic conformational changes in methionine-rich helix and the relative regulatory loop (R-loop). We further solved the structure of Cu-soaked enzyme, and found that the addition of Cu ions induced further conformational changes in the R-loop and methionine-rich helix as a result of the new Cu-binding sites on the enzyme's surface. We propose a mechanism for the enhanced laccase activity of the G304K mutant, where movements of the R-loop combined with the changes of the methionine-rich region uncover the T1 Cu site allowing greater access of the substrate. Two of the G304K double mutants showed the enhanced or decreased laccase activity, providing further evidence for the interaction between the R-loop and the methionine-rich region. The cuprous oxidase activity of these mutants was about 20% that of wild-type CueO. These structural features of the G304K mutant provide clues for designing specific substrate-binding mutants in the biotechnological applications.
WOS关键词	ESCHERICHIA-COLI ; COPPER HOMEOSTASIS ; CUPROUS OXIDASE ; FUNCTIONAL-ROLE ; SITE ; INSIGHTS ; PROTEIN ; MECHANISMS ; RESISTANCE ; RESOLUTION
WOS研究方向	Science & Technology - Other Topics
语种	英语
WOS记录号	WOS:000445336600020
内容类型	期刊论文
源URL	[http://ir.ihep.ac.cn/handle/311005/286351]
专题	高能物理研究所_多学科研究中心
作者单位	中国科学院高能物理研究所
推荐引用方式 GB/T 7714	Wang HQ,Gong Y,Dong YH,et al. Crystal structures of multicopper oxidase CueO G304K mutant: structural basis of the increased laccase activity[J]. SCIENTIFIC REPORTS,2018,8:14252.
APA	王汉潜.,宫宇.,董宇辉.,高增强.,闫玉华.,...&Wang, HQ.(2018).Crystal structures of multicopper oxidase CueO G304K mutant: structural basis of the increased laccase activity.SCIENTIFIC REPORTS,8,14252.
MLA	王汉潜,et al."Crystal structures of multicopper oxidase CueO G304K mutant: structural basis of the increased laccase activity".SCIENTIFIC REPORTS 8(2018):14252.