Chemical Reactivity of Brome Mosaic Virus Capsid Protein

• Published in: Journal of molecular biology 2012-10, Vol.423 (1), p.79-95
• Main Authors: Running, W.E., Ni, P., Kao, C.C., Reilly, J.P.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; brome mosaic virus; Bromovirus - chemistry; Capsid Proteins - chemistry; chemical modification; Hydrogen-Ion Concentration; Imidoesters - chemistry; lysine; Lysine - chemistry; mass spectrometry; Models, Molecular; Molecular Sequence Data; pH dependence; Virion - chemistry
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2012.06.031

Viral particles are biological machines that have evolved to package, protect, and deliver the viral genome into the host via regulated conformational changes of virions. We have developed a procedure to modify lysine residues with S-methylthioacetimidate across the pH range from 5.5 to 8.5. Lysine residues that are not completely modified are involved in tertiary or quaternary structural interactions, and their extent of modification can be quantified as a function of pH. This procedure was applied to the pH-dependent structural transitions of brome mosaic virus (BMV). As the reaction pH increases from 5.5 to 8.5, the average number of modified lysine residues in the BMV capsid protein increases from 6 to 12, correlating well with the known pH‐dependent swelling behavior of BMV virions. The extent of reaction of each of the capsid protein's lysine residues has been quantified at eight pH values using coupled liquid chromatography–tandem mass spectrometry. Each lysine can be assigned to one of three structural classes identified by inspection of the BMV virion crystal structure. Several lysine residues display reactivity that indicates their involvement in dynamic interactions that are not obvious in the crystal structure. The influence of several capsid protein mutants on the pH-dependent structural transition of BMV has also been investigated. Mutant H75Q exhibits an altered swelling transition accompanying solution pH increases. The H75Q capsids show increased reactivity at lysine residues 64 and 130, residues distal from the dimer interface occupied by H75, across the entire pH range. [Display omitted] ► The pH dependence of a virion’s structure is difficult to study crystallographically. ► Chemical modification of lysine residues can be quantified using mass spectrometry. ► Extent of lysine modification depends on virion structure and its pH dependence. ► Capsid protein mutants show quantitatively different labeling patterns. ► Mass‐spectrometry-detected chemical modification reliably monitors virion conformational changes.