用于核磁共振（NMR）和质谱（MS）测量的梅森- Pfizer猴病毒肉豆蔻酰化基质蛋白的表达与纯化
Expression and purification of myristoylated matrix protein of Mason-Pfizer monkey virus for NMR and MS measurements.
作者信息
Prchal Jan, Junkova Petra, Strmiskova Miroslava, Lipov Jan, Hynek Radovan, Ruml Tomas, Hrabal Richard
机构信息
Laboratory of NMR Spectroscopy, Institute of Chemical Technology, Prague, Czech Republic.
出版信息
Protein Expr Purif. 2011 Sep;79(1):122-7. doi: 10.1016/j.pep.2011.05.010. Epub 2011 May 24.
Abstract
Matrix proteins play multiple roles both in early and late stages of the viral replication cycle. Their N-terminal myristoylation is important for interaction with the host cell membrane during virus budding. We used Escherichia coli, carrying N-myristoyltransferase gene, for the expression of the myristoylated His-tagged matrix protein of Mason-Pfizer monkey virus. An efficient, single-step purification procedure eliminating all contaminating proteins including, importantly, the non-myristoylated matrix protein was designed. The comparison of NMR spectra of matrix protein with its myristoylated form revealed substantial structural changes induced by this fatty acid modification.
摘要
基质蛋白在病毒复制周期的早期和晚期都发挥着多种作用。它们的N端肉豆蔻酰化对于病毒出芽过程中与宿主细胞膜的相互作用很重要。我们使用携带N-肉豆蔻酰转移酶基因的大肠杆菌来表达肉豆蔻酰化的、带有His标签的马森- Pfizer猴病毒基质蛋白。设计了一种高效的单步纯化程序，可去除所有污染蛋白，重要的是，包括未肉豆蔻酰化的基质蛋白。基质蛋白与其肉豆蔻酰化形式的NMR光谱比较显示，这种脂肪酸修饰引起了显著的结构变化。
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