Functional Organization and Evolution of Mammalian Hexokinases: Mutations That Caused the Loss of Catalytic Activity in N-terminal Halves of Type I and Type III Isozymes

ASIA unversity > 醫學暨健康學院 > 食品營養與保健生技學系 > 期刊論文 > Item 310904400/17379

請使用永久網址來引用或連結此文件: http://asiair.asia.edu.tw/ir/handle/310904400/17379

題名:	Functional Organization and Evolution of Mammalian Hexokinases: Mutations That Caused the Loss of Catalytic Activity in N-terminal Halves of Type I and Type III Isozymes
作者:	蔡建鈞;Tsai, Henry J
貢獻者:	保健營養生技學系
關鍵詞:	hexokinase;mammalian;N-terminal half;inactivation;mutations;evolution
日期:	1999
上傳時間:	2012-11-26 02:32:14 (UTC+0)
摘要:	Mammalian hexokinases are believed to have evolved from a 100-kDa hexokinase which itself is a product of duplication and fusion of an ancestral gene encoding a 50-kDa glucose 6-phosphate-sensitive hexokinase. Type II hexokinase has been shown to possess two distinct functional active sites, one in each half, which functionally resemble the original 100-kDa hexokinase, whereas type I and III isozymes possess only one active site in the C-terminal halves. This study was conducted to identify which mutations caused the loss of catalytic activity in the N-terminal halves of type I and III isozymes. Arg 174 and Ser 447 in type I isozyme and Asp 244 in type III isozyme are speculated to be the cause, because they reside adjacent to the “catalytic” site and corresponding residues, Gly 174, Asp 447, and Gly 231, are conserved in the N-terminal half of type II isozyme as well as all other 50-kDa units that possess catalytic activity. Mutations G174R and D447S in the N-terminal half of type II isozyme reduced specific activity by approximately 79 and 57%, respectively. Therefore, neither mutation alone can account for the inactivation of the N-terminal active site in type I isozyme. Either mutation, G174R or D447S, had moderate effects on Michaelis constants, Km, for glucose and ATP · Mg2+. Intriguingly, mutation D447S introduced a novel inhibition by unchelated ATP (Ki = 68 μM ATP, competitive vs ATP · Mg2+) to the N-terminal active site of type II isozyme. Mutation G231D caused instability to type II hexokinase and near complete loss of catalytic activity (95%), suggesting that mutation G231D not only hinders catalysis at the N-terminal active site but also leads to structural instability in type II hexokinase.
關聯:	ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 369(1):149-156.
顯示於類別:	[食品營養與保健生技學系] 期刊論文

文件中的檔案:

檔案	描述	大小	格式	瀏覽次數
index.html		0Kb	HTML	271	檢視/開啟

在ASIAIR中所有的資料項目都受到原著作權保護.

資料載入中.....