Biochemical Characterization of Populus ASE Proteins
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1.
State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
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2.
University of Chinese Academy of Sciences, Beijing 100049, China
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3.
College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China
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Received Date:
2015-08-04
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Abstract
[Objective] The purpose of this study is to understand the biochemical functions of glutamine phosphoribosyl pyrophosphate amidotransferase (ASE) .[Method] By sq-PCR, protein subcellular location and catalytic activity determination of purified protein, the in vitro biochemical functions of ASE proteins were studied, and furthermore, the deletion mutant of Arabidopsis by complementation (atase2) were used to study the in vivo biological functions of poplar ASE genes. [Result] Two ASE genes were identified from Populus trichocarpa. The protein sequence analysis revealed a high sequence similarity between the two ASEs. These ASE genes were expressed in root, stem, leaf and bud tissues. The results of protein subcellular localization analysis showed that Populus ASE proteins were localized in chloroplasts. The Populus ASE genes could completely complement the function of Arabidopsis atase2 mutant. The recombinant Populus ASE proteins were expressed in E. coli and purified by Ni-affinity chromatography. Populus ASE proteins showed enzymatic activities to 5-phosphoribosyl-α-pyrophosphate. [Conclusion] It is suggested that the two Populus ASE proteins might play important roles in de novo biosynthesis of purine.
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