Monitoring the real-time kinetics of the hydrolysis reaction of guanine nucleotide-binding proteins

被引：24
作者：
Eberth, A
Dvorsky, R
Becker, CFW
Beste, A
Goody, RS
Ahmadian, MR
机构：
[1] Max Planck Inst Mol Physiol, Dept Biol Struct, D-42227 Dortmund, Germany
[2] Max Planck Inst Mol Physiol, Dept Phys Biochem, D-42227 Dortmund, Germany
[3] Ruhr Univ Bochum, Inst Physiol Chem, D-44801 Bochum, Germany
来源：
BIOLOGICAL CHEMISTRY | 2005年 / 386卷 / 11期
关键词：
fluorescence reporter; GTPase; GTPase-activating protein; GTP hydrolysis; guanine nucleotide-binding protein; rhodamine; spectroscopy;
D O I：
10.1515/BC.2005.127
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The conversion of guanosine triphosphate (GTP) to guanosine diphosphate (GDP) and inorganic phosphate (Pi) by guanine nucleotide-binding proteins (GNBPs) is a fundamental enzyme reaction in living cells that acts as an important timer in a variety of biological processes. This reaction is intrinsically slow but can be stimulated by GTPase-activating proteins (GAPs) by several orders of magnitude. In the present study, we synthesized and characterized a new fluorescent nucleotide, 2'(3')O-(N-ethylcarbamoyl-(5"-carboxytetramethylrhodamine) amide)-GTP, or tamraGTP, which is sensitive towards conformational changes of certain GNBPs induced by GTP hydrolysis. Unlike other fluorescent nucleotides, tamra-GTP allows real-time monitoring of the kinetics of the intrinsic and GAP-catalyzed GTP hydrolysis reactions of small GNBPs from the Rho family.
引用
收藏
页码：1105 / 1114
页数：10
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