Is local flexibility of M-PMV matrix protein related to  its binding affinity  with a molecular motor?


Pavel Srb1,2, Jan Lang 1,2,Jiří Vlach2, Marian Grocký 1, Jan Prchal3, Jan Lipov3, Tomáš Ruml3,4 and Richard Hrabal2


1Department of Low Temperature Physics, Faculty of Mathematics and Physics, Charles University, V Holešovičkách 2, 180 00 Prague 8, Czech Republic

2Laboratory of NMR Spectroscopy and

3Department of Biochemistry and Microbiology

Institute of Chemical Technology, Technická 5, 166 28 Prague 6, Czech Republic

4Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech republic, Flemingovo nám. 2, 166 10, Prague 6, Czech Republic


Mason-Pfizer monkey virus (M-PMV) belongs to the genus of betaretroviruses in which the matrix protein (MA) plays the essential role in certain stages of their life cycle (e.g. in assembly, transport and budding of new viral particles). Several single or double point mutants are known to cause dramatic changes in the virus life cycle [1]. In particular, the single point mutation R55F in MA redirects the assembly of the viral capsid to the plasma membrane instead of to cytoplasm, which is the place of assembly of the wild type form (WT).

The NMR assignments and three-dimensional structures of WT and R55F have been solved recently by our group [2]. Both structures are composed of four a-helices,  however the relative orientation of the N-terminal domain   (helices 1 and 2) with respect to the C-terminal one  (helices 3 and 4) is different.  Such reorientation causes different accessibility of Cytoplasm Targetting/Retention Signal (CTRS) sequence for the interaction with a molecular motor dynein . It is expected that the degree of flexibility of the CTRS sequence will affect binding affinity with the Tctex-1 which is one of the light chains of dynein. 

Therefore, we carried out a detailed motional analysis of the loop connecting helices 2 and 3, the least structured  part of CTRS. Relaxation parameters of amide 15N and carbonyl 13C nuclei have been studied by NMR spectroscopy on  uniformly labelled (15N and 15N/13C) samples. Additionally, MD trajectory analysis have provided a complementary view on the mobility of CTRS sequence which is crucial for understanding the interation of Gag with dynein.

1.      S. S. Rhee and E. Hunter, Cell, 1990, 63, 77–86

2.       J. Vlach, J. Lipov, V.Veverka, M.Rumlova, T. Ruml, R.Hrabal, J.Biomol. NMR, 2005, 4, 381-2