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.

Laboratory of NMR Spectroscopy Institute of Chemical Technology Technická 5 16628 Prague Czech Republic; Department of Biochemistry and Microbiology Institute of Chemical Technology Technická 5 16628 Prague Czech Republic; Institute of Organic Chemistry and Biochemistry Academy of Sciences of the Czech Republic v v i IOCB and Gilead Research Center Flemingovo nám 2 16610 Prague Czech Republic

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Myristoylation drives dimerization of matrix protein from mouse mammary tumor virus