Binding of ATP to the fructose-2,6-bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase leads to activation of its 6-phosphofructo-2-kinase

	Binding of ATP to the fructose-2,6-bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase leads to activation of its 6-phosphofructo-2-kinase
	Yang, QH; Zhu, Z; Dong, MQ; Ling, S; Wu, CL; Li, L
刊名	JOURNAL OF BIOLOGICAL CHEMISTRY
	2001
卷号	276 期号:27 页码:24608-24613
通讯作者	Li, L (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, State Key Lab Mol Biol, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,
英文摘要	To understand the mechanism by which the activity of the 6-phosphofructo-2-kinase (6PF-2K) of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase is stimulated by its substrate ATP, we studied two mutants of the enzyme. Mutation of either Arg-279, the penultimate basic residue within the Walker A nucleotide-binding fold in the bisphosphatase domain, or Arg-359 to Ala eliminated the activation of the chicken 6PF-2K by ATP, Binding analysis by fluorescence spectroscopy using 2 ' (3 ')-O-(N-methylanthraniloyl)-ATP revealed that the kinase domains of these two mutants, unlike that of the wild type enzyme, showed no cooperativity in ATP binding and that the mutant enzymes possess only the high affinity ATP binding site, suggesting that the ATP binding site on the bisphosphatase domain represents the low affinity site. This conclusion was supported by the result that the affinity of ATP for the isolated bisphosphatase domain is similar to that for the low affinity site in the wild type enzyme. In addition, we found that the 6PP-2K of a chimeric enzyme, in which the last 25 residues of chicken enzyme were replaced with those of the rat enzyme, could not be activated by ATP, despite the fact that the ATP-binding properties of this chimeric enzyme were not different from those of the wild type chicken enzyme, These results demonstrate that activation of the chicken 6PF-2K by ATP may result from allosteric binding of ATP to the bisphosphatase domain where residues Arg-279 and Arg-359 are critically involved and require specific C-terminal sequences.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	SITE-DIRECTED MUTAGENESIS ; RAT-LIVER ; BIFUNCTIONAL ENZYME ; 2-KINASE DOMAIN ; CRYSTAL-STRUCTURE ; FRUCTOSE 6-PHOSPHATE,2-KINASE ; ESCHERICHIA-COLI ; BASIC RESIDUES ; MECHANISM ; 2,6-BISPHOSPHATASE
收录类别	SCI
语种	英语
WOS记录号	WOS:000169800700027
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2721]
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	Yang, QH,Zhu, Z,Dong, MQ,et al. Binding of ATP to the fructose-2,6-bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase leads to activation of its 6-phosphofructo-2-kinase[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2001,276(27):24608-24613.
APA	Yang, QH,Zhu, Z,Dong, MQ,Ling, S,Wu, CL,&Li, L.(2001).Binding of ATP to the fructose-2,6-bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase leads to activation of its 6-phosphofructo-2-kinase.JOURNAL OF BIOLOGICAL CHEMISTRY,276(27),24608-24613.
MLA	Yang, QH,et al."Binding of ATP to the fructose-2,6-bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase leads to activation of its 6-phosphofructo-2-kinase".JOURNAL OF BIOLOGICAL CHEMISTRY 276.27(2001):24608-24613.