Weak protein-protein interactions are sufficient to drive assembly of hepatitis B virus capsids.
Hepatitis B virus (HBV) is an enveloped DNA virus with a spherical capsid (or core). The capsid is constructed from 120 copies of the homodimeric capsid protein arranged with T = 4 icosahedral symmetry. We examined in vitro assembly of purified E. coli expressed HBV capsid protein. After equilibration, concentrations of capsid and dimer were evaluated by size exclusion chromatography. The extent of assembly increased as temperature and ionic strength increased. The concentration dependence of capsid assembly conformed to the equilibrium expression: K(capsid) = [capsid]/[dimer](120). Given the known geometry for HBV capsids and dimers, the per capsid assembly energy was partitioned into energy per subunit-subunit contact. We were able to make three major conclusions. (i) Weak interactions (from -2.9 kcal/mol at 21 degrees C in low salt to -4.4 kcal/mol at 37 degrees C in high salt) at each intersubunit contact result in a globally stable capsid; weak intersubunit interactions may be the basis for the phenomenon of capsid breathing. (ii) HBV assembly is characterized by positive enthalpy and entropy. The reaction is entropy-driven, consistent with the largely hydrophobic contacts found in the crystal structure. (iii) Increasing NaCl concentration increases the magnitude of free energy, enthalpy, and entropy, as if ionic strength were increasing the amount of hydrophobic surface buried by assembly. This last point leads us to suggest that salt acts by inducing a conformational change in the dimer from an assembly-inactive form to an assembly-active form. This model of conformational change linked to assembly is consistent with immunological differences between dimer and capsid.
Ceres
Pablo
P
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA.
Zlotnick
Adam
A
D036022:Capsid Proteins / Q000737:chemistry* / Q000502:physiology*; D019281:Dimerization; D006511:Hepatitis B Antigens / Q000737:chemistry* / Q000502:physiology*; D006515:Hepatitis B virus / Q000737:chemistry* / Q000502:physiology*; D007700:Kinetics; D011487:Protein Conformation; D012492:Salts; D012965:Sodium Chloride / Q000737:chemistry; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D019065:Virus Assembly
D036022:Capsid Proteins; D006511:Hepatitis B Antigens; D012492:Salts; D012965:Sodium Chloride