Structural Insights into Cellulolytic and Chitinolytic Enzymes Revealing Crucial Residues of Insect beta-N-acetyl-D-hexosaminidase

doi:10.1371/journal.pone.0052225

CORC > 上海药物研究所 > 中国科学院上海药物研究所 > 药理学第三研究室

	Structural Insights into Cellulolytic and Chitinolytic Enzymes Revealing Crucial Residues of Insect beta-N-acetyl-D-hexosaminidase
	Liu, Tian 4; Zhou, Yong 1; Chen, Lei 4; Chen, Wei 4; Liu, Lin 4; Shen, Xu2 ; Zhang, Wenqing 3; Zhang, Jianzhen 5; Yang, Qing 4
刊名	PLOS ONE
	2012-12-27
卷号	7 期号:12
ISSN号	1932-6203
DOI	10.1371/journal.pone.0052225
文献子类	Article
英文摘要	The chemical similarity of cellulose and chitin supports the idea that their corresponding hydrolytic enzymes would bind beta-1,4-linked glucose residues in a similar manner. A structural and mutational analysis was performed for the plant cellulolytic enzyme BGlu1 from Oryza sativa and the insect chitinolytic enzyme OfHex1 from Ostrinia furnacalis. Although BGlu1 shows little amino-acid sequence or topological similarity with OfHex1, three residues (Trp(490), Glu(328), Val(327) in OfHex1, and Trp(358), Tyr(131) and Ile(179) in BGlu1) were identified as being conserved in the +1 sugar binding site. OfHex1 Glu(328) together with Trp(490) was confirmed to be necessary for substrate binding. The mutant E328A exhibited a 8-fold increment in K-m for (GlcNAc) 2 and a 42-fold increment in K-i for TMG-chitotriomycin. A crystal structure of E328A in complex with TMG-chitotriomycin was resolved at 2.5 angstrom, revealing the obvious conformational changes of the catalytic residues (Glu(368) and Asp(367)) and the absence of the hydrogen bond between E328A and the C3-OH of the +1 sugar. V327G exhibited the same activity as the wild-type, but acquired the ability to efficiently hydrolyse beta-1,2-linked GlcNAc in contrast to the wild-type. Thus, Glu(328) and Val(327) were identified as important for substrate-binding and as glycosidic-bond determinants. A structure-based sequence alignment confirmed the spatial conservation of these three residues in most plant cellulolytic, insect and bacterial chitinolytic enzymes.
资助项目	National Key Project for Basic Research[2010CB126100] ; National Natural Science Foundation of China[31070715] ; National Natural Science Foundation of China[31101671] ; National High Technology Research and Development Program of China[2011AA10A204] ; National Key Technology RD Program[2011BAE06B05] ; Fundamental Research Funds for the Central Universities[DUT11ZD113] ; Fundamental Research Funds for the Central Universities[DUT11RC(3)73]
WOS关键词	OSTRINIA-FURNACALIS ; RICE BGLU1 ; CATALYTIC MECHANISM ; D-GLUCOSIDASE ; PURIFICATION ; SEQUENCE ; CHITOBIASE ; PROTEINS ; BINDING ; GENOME
WOS研究方向	Science & Technology - Other Topics
语种	英语
出版者	PUBLIC LIBRARY SCIENCE
WOS记录号	WOS:000312829100032
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/277837]
专题	药理学第三研究室中科院受体结构与功能重点实验室新药研究国家重点实验室
通讯作者	Yang, Qing
作者单位	1.Dalian Univ Technol, Sch Software Technol, Dalian, Peoples R China; 2.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 200031, Peoples R China; 3.Sun Yat Sen Univ, Sch Life Sci, State Key Lab Biocontrol, Guangzhou 510275, Guangdong, Peoples R China; 4.Dalian Univ Technol, Sch Life Sci & Biotechnol, Dalian, Peoples R China; 5.Shanxi Univ, Res Inst Appl Biol, Taiyuan, Peoples R China
推荐引用方式 GB/T 7714	Liu, Tian,Zhou, Yong,Chen, Lei,et al. Structural Insights into Cellulolytic and Chitinolytic Enzymes Revealing Crucial Residues of Insect beta-N-acetyl-D-hexosaminidase[J]. PLOS ONE,2012,7(12).
APA	Liu, Tian.,Zhou, Yong.,Chen, Lei.,Chen, Wei.,Liu, Lin.,...&Yang, Qing.(2012).Structural Insights into Cellulolytic and Chitinolytic Enzymes Revealing Crucial Residues of Insect beta-N-acetyl-D-hexosaminidase.PLOS ONE,7(12).
MLA	Liu, Tian,et al."Structural Insights into Cellulolytic and Chitinolytic Enzymes Revealing Crucial Residues of Insect beta-N-acetyl-D-hexosaminidase".PLOS ONE 7.12(2012).