One-step separation of myristoylated and nonmyristoylated retroviral matrix proteins
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
24056256
DOI
10.1016/j.pep.2013.09.003
PII: S1046-5928(13)00181-2
Knihovny.cz E-resources
- Keywords
- Matrix protein, Mouse mammary tumor virus, Murine leukemia virus, Myristoylation, N-myristoyltransferase, Retrovirus,
- MeSH
- Chromatography, Affinity MeSH
- Cloning, Molecular MeSH
- Myristic Acid chemistry metabolism MeSH
- Mice MeSH
- Nuclear Magnetic Resonance, Biomolecular MeSH
- Recombinant Proteins chemistry genetics isolation & purification metabolism MeSH
- Retroviridae Proteins chemistry genetics isolation & purification metabolism MeSH
- Retroviridae Infections virology MeSH
- Leukemia Virus, Murine chemistry genetics metabolism MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Myristic Acid MeSH
- Recombinant Proteins MeSH
- Retroviridae Proteins MeSH
N-terminal myristoylation of retroviral matrix proteins is essential for the targeting of the Gag polyproteins to the plasma membrane. To investigate the effect of the myristoylation on the structure and membrane binding ability of the matrix proteins, it is necessary to prepare their myristoylated forms. We present purification of myristoylated matrix proteins of the mouse mammary tumor virus and murine leukemia virus, two morphogenetically distinct retroviruses. The proteins were expressed in Escherichia coli coexpressing a yeast N-myristoyltransferase. This E. coli expression system yielded a mixture of myristoylated and nonmyristoylated matrix proteins. We established efficient one-step metal affinity purification that enabled to obtain pure myristoylated matrix proteins suitable for structural and functional studies.
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