Preparation and structure determination of T41I/T78I mutant of the M-PMV matrix protein

Tomas Kroupa^1,2, Jan Prchal^1,2, Richard Hrabal²

¹Department of Biochemistry and Microbiology, Institute of Chemical Technology,Technická 5, 166 28 Prague 6, Czech Republic

²Laboratory of NMR Spectroscopy, Institute of Chemical Technology, Technická 5, 166 28 Prague 6, Czech Republic

richard.hrabal@vscht.cz

Mason-Pfizer monkey virus (M-PMV) belongs to betaretroviruses and is widely used as a model organism for studies of the late phase of the life cycle of retroviruses. Matrix protein which is naturally modified by the rest of the myristic acid on the N-terminus, plays key role in the transportation of the immature retroviral particles to budding sites and is responsible for an interaction with the plasma membrane. Mutation T41I/T78I in the matrix protein causes either a change in the hydrophobicity of the cavity in which the myristoyl is nested or a change in an interaction site of the matrix protein with the plasma membrane. The outcome is that the immature viral particles can't bud from the infected cell.

Our work focused on the preparation of the myristoylated T41I/T78I mutant of the matrix protein of M-PMV. The mutant protein was analyzed by the NMR spectroscopy and the 3D structure was determined. The liposome binding assay was used to study an interaction with the plasma membrane.