通过N-肉豆蔻酰甘氨酸的氮杂环丙烷衍生物的反相高效液相色谱法鉴定N-肉豆蔻酰化蛋白。
Identification of N-myristoylated proteins by reverse-phase high performance liquid chromatography of an azlactone derivative of N-myristoylglycine.
作者信息
Goddard C, Felsted R L
机构信息
Laboratory of Biological Chemistry, National Cancer Institute, Bethesda, MD 20892.
出版信息
Biochem J. 1988 Aug 1;253(3):839-43. doi: 10.1042/bj2530839.
Abstract
A method is presented for the identification of N-myristoylated proteins. N-Myristoyl transferases have an absolute requirement for a free N-terminal glycine. N-Myristoylglycine is released upon mild acid hydrolysis of myristoylated peptides and proteins and its derivitization to a p-nitrobenzylazlactone with subsequent analysis by reverse phase h.p.l.c. enabled its detection to pmol levels. This facilitated the identification of N-terminal myristate in nmol quantities of purified proteins. Using this method we demonstrate that the alpha-subunit of the GTP-binding protein G0 is N-terminally myristoylated.
摘要
本文介绍了一种鉴定N-肉豆蔻酰化蛋白的方法。N-肉豆蔻酰转移酶对游离的N端甘氨酸有绝对需求。肉豆蔻酰化的肽和蛋白质经温和酸水解后会释放出N-肉豆蔻酰甘氨酸，将其衍生化为对硝基苄基氮杂环丁烷酮，随后通过反相高效液相色谱分析，可将其检测至皮摩尔水平。这有助于鉴定纳摩尔量纯化蛋白中的N端肉豆蔻酸。使用该方法，我们证明了GTP结合蛋白G0的α亚基在N端被肉豆蔻酰化。
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