胃饥饿素水解酶活性的放射性测定及3H-胃饥饿素在小鼠组织中的分布。
Radiometric assay of ghrelin hydrolase activity and 3H-ghrelin distribution into mouse tissues.
作者信息
Chen Vicky Ping, Gao Yang, Geng Liyi, Brimijoin Stephen
机构信息
Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA; Kogod Center on Aging, Mayo Clinic, Rochester, MN 55905, USA.
出版信息
Biochem Pharmacol. 2015 Dec 15;98(4):732-9. doi: 10.1016/j.bcp.2015.10.016. Epub 2015 Oct 26.
Abstract
A high-throughput radiometric assay was developed to characterize enzymatic hydrolysis of ghrelin and to track the peptide's fate in vivo. The assay is based on solvent partitioning of [(3)H]-octanoic acid liberated from [(3)H]-octanoyl ghrelin during enzymatic hydrolysis. This simple and cost-effective method facilitates kinetic analysis of ghrelin hydrolase activity of native and mutated butyrylcholinesterases or carboxylesterases from multiple species. In addition, the assay's high sensitivity facilitates ready evaluation of ghrelin's pharmacokinetics and tissue distribution in mice after i.v. bolus administration of radiolabeled peptide.
摘要
开发了一种高通量放射性测定法，以表征胃饥饿素的酶促水解，并追踪该肽在体内的去向。该测定法基于酶促水解过程中从[(3)H]-辛酰胃饥饿素释放的[(3)H]-辛酸的溶剂分配。这种简单且经济高效的方法有助于对来自多个物种的天然和突变丁酰胆碱酯酶或羧酸酯酶的胃饥饿素水解酶活性进行动力学分析。此外，该测定法的高灵敏度有助于在静脉推注放射性标记肽后对小鼠体内胃饥饿素的药代动力学和组织分布进行快速评估。
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本文引用的文献
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Gene transfer of mutant mouse cholinesterase provides high lifetime expression and reduced cocaine responses with no evident toxicity.基因转移的突变型小鼠胆碱酯酶提供了高终身表达和减少可卡因的反应，没有明显的毒性。
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