Nonmyristoylated matrix protein from the Mason-Pfizer monkey virus forms oligomers.
We studied the oligomeric properties of betaretroviral nonmyristoylated matrix protein (MA) and its R55F mutant from the Mason-Pfizer monkey virus in solution by means of chemical crosslinking and NMR spectroscopy. By analyzing crosslinked products and using concentration-dependent NMR chemical shift mapping, we have proven that the wild-type (WT) MA forms oligomers in solution. Conversely, no oligomerization was observed for the R55F mutant. Structural comparison of MAs explained their different behaviors in solution, concluding that the key residues involved in intermonomeric interaction are exposed in the WT MA but buried in the mutant, preventing the oligomerization of R55F. The final model of oligomerization of the WT MA was derived by concerted use of chemical shift mapping and diffusion-ordered spectroscopy measured on a set of protein samples with varying concentrations. We found that the Mason-Pfizer monkey virus WT MA exists in a monomer-dimer-trimer equilibrium in solution, with the corresponding dissociation constants of 2.3 and 0.24 mM, respectively. Structures of the oligomers calculated with HADDOCK software are closely related to the structures of other retroviral MA trimers.
Vlach
Jirí
J
Institute of Chemical Technology, Prague, Czech Republic.
Srb
Pavel
P
Prchal
Jan
J
Grocký
Marián
M
Lang
Jan
J
Ruml
Tomás
T
Hrabal
Richard
R
D000595:Amino Acid Sequence; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004058:Diffusion; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D016093:Mason-Pfizer monkey virus / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry; D019814:Myristic Acid / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D055503:Protein Multimerization / Q000187:drug effects; D020836:Protein Structure, Quaternary; D016415:Sequence Alignment; D014763:Viral Matrix Proteins / Q000737:chemistry*
D003432:Cross-Linking Reagents; D050505:Mutant Proteins; D014763:Viral Matrix Proteins; D019814:Myristic Acid