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Effects of removing a conserved disulfide bond on the biological characteristics of rat lipocalin-type prostaglandin D synthase

文献类型：期刊论文

作者Liu, Jiafu2; Guo, Chenyun1; Yao, Yihe1; Lin, Donghai1,2
刊名BIOCHIMIE
出版日期2008-11
卷号90期号:11-12页码:1637-1646
关键词Lipocalin-type prostaglandin D synthase Variant Disulfide bond NMR
ISSN号0300-9084
DOI10.1016/j.biochi.2008.06.003
文献子类Article
英文摘要Three cysteine residues, Cys(65), Cys(89), and Cys(186) in lipocalin-type prostaglandin D synthase (L-PGDS), are conserved among all species and the disulfide bond between Cys(89) and Cys(186) is highly conserved among most, but not all, lipocalins. In this study, four rat L-PGDS variants were constructed by site-directed mutagenesis, and the conserved disulfide bond in several variants was removed by substituting cysteine with alanine. The effects of removing this disulfide bond on their biological characteristics were investigated. The NMR experiments indicated that the removal of disulfide did not change their conformations significantly. However, both thermal-induced and urea-induced unfolding experiments showed that the stabilities of enzymes without the disulfide bond decreased significantly. Moreover, the ligand-binding affinities of these variants were assessed by fluorescence experiments. Dissociation constants (K-d) of 0.668, 0.689, 0.543 and 0.571 mu M were obtained for ANS binding to wild-type rat L-PGDS, C(65)A, C(186)A, and C(89,186)A variants, respectively, and 71.2 and 62.3 nM for retinoic acid binding to wild-type rat L-PGDS and the C(186)A variant, respectively. These results suggested that the removal of the disulfide bond slightly increased the affinities for ligand binding by changing the hydrophobic regions. This study may offer valuable information for further studies on other rat lipocalins. (c) 2008 Elsevier Masson SAS. All rights reserved.
WOS关键词MOLTEN-GLOBULE ; FLUORESCENT-PROBE ; BINDING PROTEINS ; LIGAND-BINDING ; EXPRESSION ; ACID ; IDENTIFICATION ; INTERMEDIATE ; SULFONATE ; STATES
WOS研究方向Biochemistry & Molecular Biology
语种英语
WOS记录号WOS:000261567400004
出版者ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
源URL[http://119.78.100.183/handle/2S10ELR8/279442]  
专题分析化学研究室
通讯作者Liu, Jiafu
作者单位1.Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai Inst Mat Med, Nanjing 201203, Peoples R China
2.China Pharmaceut Univ, Sch Life Sci & Technol, Nanjing 210009, Peoples R China;
推荐引用方式
GB/T 7714		 Liu, Jiafu,Guo, Chenyun,Yao, Yihe,et al. Effects of removing a conserved disulfide bond on the biological characteristics of rat lipocalin-type prostaglandin D synthase[J]. BIOCHIMIE,2008,90(11-12):1637-1646.
APA Liu, Jiafu,Guo, Chenyun,Yao, Yihe,&Lin, Donghai.(2008).Effects of removing a conserved disulfide bond on the biological characteristics of rat lipocalin-type prostaglandin D synthase.BIOCHIMIE,90(11-12),1637-1646.
MLA Liu, Jiafu,et al."Effects of removing a conserved disulfide bond on the biological characteristics of rat lipocalin-type prostaglandin D synthase".BIOCHIMIE 90.11-12(2008):1637-1646.

入库方式： OAI收割

来源：上海药物研究所

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