Leptospira interrogans encodes an ROK family glucokinase involved in a cryptic glucose utilization pathway

	Leptospira interrogans encodes an ROK family glucokinase involved in a cryptic glucose utilization pathway
	Zhang, Q; Zhang, YY; Zhong, Y; Ma, J; Peng, NQ; Cao, XJ; Yang, C; Zeng, R; Guo, XK; Zhao, GP
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2011
卷号	43 期号:8 页码:618-629
关键词	Leptospira interrogans glucokinase glucose utilization
通讯作者	Zhao, GP (reprint author), Chinese Acad Sci, Key Lab Synthet Biol, Inst Plant Physiol & Ecol, Shanghai Inst Biol Sci, Shanghai 200032, Peoples R China.,gpzhao@sibs.ac.cn
英文摘要	Although Leptospira interrogans is unable to utilize glucose as its carbon/energy source, the LA_1437 gene of L. interrogans serovar Lai potentially encodes a group III glucokinase (GLK). The L. interrogans GLK (LiGLK) heterogeneously expressed in Escherichia coli was purified and proved to be a homodimeric enzyme with its specific activity of 12.3 +/- 0.6 U/mg. protein determined under an improved assay condition (pH 9.0, 50 degrees C), 7.5-fold higher than that assayed under the previously used condition (pH 7.3, 25 degrees C). The improved sensitivity allowed us to detect this enzymatic activity of (5.0 +/- 0.6)x10(-3) U/mg. protein in the crude extract of L. interrogans serovar Lai cultured in standard Ellinghausen-McCullough-Johnson-Harris medium. The k(cat) and K(m) values for D-glucose and ATP were similar to those of other group III GLKs, although the K(m) value for ATP was slightly higher. Site-directed mutagenesis analysis targeting the conserved amino acid residues in the potential ATP-binding motif hinted that a proper array of Gly residues in the motif might be important for maintaining the conformation that was essential for its function. Gene expression profiling and quantitative proteomic data mining provided preliminary evidence for the absence of efficient systems involved in glucose transport and glycolysis that might account for the failure of glucose utilization in L. interrogans.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	ATP-DEPENDENT GLUCOKINASE ; HYPERTHERMOPHILIC ARCHAEON ; BIOCHEMICAL-CHARACTERIZATION ; THERMOTOGA-MARITIMA ; BACILLUS-SUBTILIS ; ESCHERICHIA-COLI ; SUGAR KINASES ; PROTEIN ; EXPRESSION ; CLONING
收录类别	SCI
语种	英语
WOS记录号	WOS:000293299600006
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/838]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Zhang, Q,Zhang, YY,Zhong, Y,et al. Leptospira interrogans encodes an ROK family glucokinase involved in a cryptic glucose utilization pathway[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2011,43(8):618-629.
APA	Zhang, Q.,Zhang, YY.,Zhong, Y.,Ma, J.,Peng, NQ.,...&Zhao, GP.(2011).Leptospira interrogans encodes an ROK family glucokinase involved in a cryptic glucose utilization pathway.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,43(8),618-629.
MLA	Zhang, Q,et al."Leptospira interrogans encodes an ROK family glucokinase involved in a cryptic glucose utilization pathway".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 43.8(2011):618-629.