Substrate binding of a GH5 endoglucanase from the ruminal fungus Piromyces rhizinflata

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	Substrate binding of a GH5 endoglucanase from the ruminal fungus Piromyces rhizinflata
	Tseng, Chih-Wen 1; Ko, Tzu-Ping 2; Guo, Rey-Ting 3; Huang, Jian-Wen 1; Wang, Hao-Ching 2; Huang, Chun-Hsiang 3; Cheng, Ya-Shan 1; Wang, Andrew H. -J.2; Liu, Je-Ruei 1,4,5
刊名	ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS
	2011-10-01
卷号	67 期号:Part 10 页码:1189-1194
英文摘要	The endoglucanase EglA from Piromyces rhizinflata found in cattle stomach belongs to the GH5 family of glycoside hydrolases. The crystal structure of the catalytic domain of EglA shows the (beta/alpha)(8)-barrel fold typical of GH5 enzymes. Adjacent to the active site of EglA, a loop containing a disulfide bond not found in other similar structures may participate in substrate binding. Because the active site was blocked by the N-terminal His tag of a neighbouring protein molecule in the crystal, enzyme-substrate complexes could not be obtained by soaking but were prepared by cocrystallization. The E154A mutant structure with a cellotriose bound to the -3, -2 and -1 subsites shows an extensive hydrogen-bonding network between the enzyme and the substrate, along with a stacking interaction between Trp44 and the -3 sugar. A possible dimer was observed in the crystal structure, but retention of activity in the E242A mutant suggested that the enzyme probably does not function as a dimer in solution. On the other hand, the first 100 amino acids encoded by the original cDNA fragment are very similar to those in the last third of the (beta/alpha)(8)-barrel fold, indicating that EglA comprises at least two catalytic domains acting in tandem.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
类目[WOS]	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biophysics ; Crystallography
研究领域[WOS]	Biochemistry & Molecular Biology ; Biophysics ; Crystallography
关键词[WOS]	CRYSTAL-STRUCTURE ; CATALYTIC DOMAIN ; ACTIVE-SITE ; CELLULASE ; SUITE ; NUCLEOPHILE ; REFINEMENT ; ENZYME ; CEL5A
收录类别	SCI
语种	英语
WOS记录号	WOS:000296785700008
公开日期	2011-10-17
内容类型	期刊论文
源URL	[http://localhost/handle/0/172]
专题	天津工业生物技术研究所_结构生物学与蛋白酶学实验室郭瑞庭_期刊论文
作者单位	1.Natl Taiwan Univ, Inst Biotechnol, Taipei 10617, Taiwan 2.Acad Sinica, Inst Biol Chem, Taipei 11529, Taiwan 3.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Ind Enzymes Natl Engn Lab, Tianjin 300308, Peoples R China 4.Natl Taiwan Univ, Dept Anim Sci & Technol, Taipei 10617, Taiwan 5.Acad Sinica, Agr Biotechnol Res Ctr, Taipei 11529, Taiwan
推荐引用方式 GB/T 7714	Tseng, Chih-Wen,Ko, Tzu-Ping,Guo, Rey-Ting,et al. Substrate binding of a GH5 endoglucanase from the ruminal fungus Piromyces rhizinflata[J]. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS,2011,67(Part 10):1189-1194.
APA	Tseng, Chih-Wen.,Ko, Tzu-Ping.,Guo, Rey-Ting.,Huang, Jian-Wen.,Wang, Hao-Ching.,...&Liu, Je-Ruei.(2011).Substrate binding of a GH5 endoglucanase from the ruminal fungus Piromyces rhizinflata.ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS,67(Part 10),1189-1194.
MLA	Tseng, Chih-Wen,et al."Substrate binding of a GH5 endoglucanase from the ruminal fungus Piromyces rhizinflata".ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS 67.Part 10(2011):1189-1194.