Balanced electrostatic and structural forces guide the large conformational change associated with maturation of T = 4 virus.
Nudaurelia capensis omega virus has a well-characterized T = 4 capsid that undergoes a pH-dependent large conformational changes (LCC) and associated auto-catalytic cleavage of the subunit. We examined previously the particle size at different pH values and showed that maturation occurred at pH 5.5. We now characterized the LCC with time-resolved small-angle x-ray scattering and showed that there were three kinetic stages initiated with an incremental drop in pH: 1), a rapid (<10 ms) collapse to an incrementally smaller particle; 2), a continuous size reduction over the next 5 s; and 3), a smaller final transition occurring in 2-3 min. Equilibrium measurements similar to those reported previously, but now more precise, showed that the particle dimension between pH 5.5 and 5 requires the autocatalytic cleavage to achieve its final compact size. A balance of electrostatic and structural forces shapes the energy landscape of the LCC with the latter requiring annealing of portions of the subunit. Equilibrium experiments showed that many intermediate states could be populated with a homogeneous ensemble of particles by carefully controlling the pH. A titration curve for the LCC was generated that showed that the virtual pK(a) (i.e., the composite of all titratable residues that contribute to the LCC) is 5.8.
Matsui
Tsutomu
T
Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, USA.
Tsuruta
Hiro
H
Johnson
John E
JE
D001703:Biophysics / Q000379:methods*; D002213:Capsid / Q000737:chemistry / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008968:Molecular Conformation; D009154:Mutation; D016011:Normal Distribution; D011487:Protein Conformation; D012542:Scattering, Radiation; D012984:Software; D055672:Static Electricity; D013997:Time Factors; D014780:Viruses / Q000378:metabolism; D014965:X-Rays
D036022:Capsid Proteins; C092202:coat protein, Nudaurelia capensis omega virus