Document Detail


Expression, purification, and characterization of recombinant Saccharomyces cerevisiae adenosine kinase.
MedLine Citation:
PMID:  12883639     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Adenosine kinase (AK), a key enzyme in the regulation of the cellular concentrations of adenosine (A), is an important physiological effector of many cells and tissues. In this article, we reported that ak, which encoded adenosine kinase, was cloned from Saccharomyces cerevisiae, sequenced, and overexpressed in E. coli using the pET16b expression system, and the recombinant protein was purified to apparent homogeneity using conventional protein purification techniques. Kinetic analysis of S. cerevisiae AK revealed K(m) values of (3.5+/-0.2) micromol/L for adenosine and (100.0+/-11.0) micromol/L for ATP, with k(cat) of (1530+/-20) min(-1) for adenosine and (1448+/-25) min(-1) for ATP. The determination of the K(m) value for other nucleosides and deoxynucleoside indicated that the nucleoside specificity of this enzyme from yeast was quite high.
Authors:
Xiao-Bing Lu; Hai-Zhen Wu; Jiang Ye; Yi Fan; Hui-Zhan Zhang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica     Volume:  35     ISSN:  0582-9879     ISO Abbreviation:  Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-07-28     Completed Date:  2003-12-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  20730160R     Medline TA:  Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)     Country:  China    
Other Details:
Languages:  eng     Pagination:  666-70     Citation Subset:  IM    
Affiliation:
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. huizhzh@online.sh.cn
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MeSH Terms
Descriptor/Qualifier:
Adenosine Kinase / genetics*,  isolation & purification,  metabolism
Escherichia coli / genetics
Kinetics
Recombinant Proteins / biosynthesis,  isolation & purification
Saccharomyces cerevisiae / enzymology*
Saccharomyces cerevisiae Proteins / genetics*
Chemical
Reg. No./Substance:
0/Recombinant Proteins; 0/Saccharomyces cerevisiae Proteins; EC 2.7.1.20/Adenosine Kinase

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