Monitoring Assembly of Virus Capsids with Nanofluidic Devices.
Virus assembly is a coordinated process in which typically hundreds of subunits react to form complex, symmetric particles. We use resistive-pulse sensing to characterize the assembly of hepatitis B virus core protein dimers into T = 3 and T = 4 icosahedral capsids. This technique counts and sizes intermediates and capsids in real time, with single-particle sensitivity, and at biologically relevant concentrations. Other methods are not able to produce comparable real-time, single-particle observations of assembly reactions below, near, and above the pseudocritical dimer concentration, at which the dimer and capsid concentrations are approximately equal. Assembly reactions across a range of dimer concentrations reveal three distinct patterns. At dimer concentrations as low as 50 nM, well below the pseudocritical dimer concentration of 0.5 μM, we observe a switch in the ratio of T = 3 to T = 4 capsids, which increases with decreasing dimer concentration. Far above the pseudocritical dimer concentration, kinetically trapped, incomplete T = 4 particles assemble rapidly, then slowly anneal into T = 4 capsids. At all dimer concentrations tested, T = 3 capsids form more rapidly than T = 4 capsids, suggesting distinct pathways for the two forms.
Department of Chemistry and ‡Department of Molecular and Cellular Biochemistry, Indiana University , Bloomington, Indiana 47405, United States.
Zachary D
ZD
Harms
Department of Chemistry and ‡Department of Molecular and Cellular Biochemistry, Indiana University , Bloomington, Indiana 47405, United States.
Lisa
L
Selzer
Department of Chemistry and ‡Department of Molecular and Cellular Biochemistry, Indiana University , Bloomington, Indiana 47405, United States.
Adam
A
Zlotnick
Department of Chemistry and ‡Department of Molecular and Cellular Biochemistry, Indiana University , Bloomington, Indiana 47405, United States.
Stephen C
SC
Jacobson
D002213:Capsid / Q000737:chemistry / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry* / Q000378:metabolism; D006515:Hepatitis B virus / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D055503:Protein Multimerization; D014771:Virion / Q000737:chemistry / Q000378:metabolism; D019065:Virus Assembly
D036022:Capsid Proteins
hepatitis B virus; in-plane nanochannel; nanofluidics; resistive-pulse sensing; self-assembly; single-particle counting