A molecular breadboard: Removal and replacement of subunits in a hepatitis B virus capsid |
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| Authors: | Lye Siang Lee Nicholas Brunk Daniel G Haywood David Keifer Elizabeth Pierson Panagiotis Kondylis Joseph Che‐Yen Wang Stephen C Jacobson Martin F Jarrold Adam Zlotnick |
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| Institution: | 1. Molecular and Cellular Biochemistry Department, Indiana University, Bloomington, Indiana;2. Intelligent Systems Engineering Department, Bloomington, Indiana;3. Department of Chemistry, Indiana University, Bloomington, Indiana |
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| Abstract: | Hepatitis B virus (HBV) core protein is a model system for studying assembly and disassembly of icosahedral structures. Controlling disassembly will allow re‐engineering the 120 subunit HBV capsid, making it a molecular breadboard. We examined removal of subunits from partially crosslinked capsids to form stable incomplete particles. To characterize incomplete capsids, we used two single molecule techniques, resistive‐pulse sensing and charge detection mass spectrometry. We expected to find a binomial distribution of capsid fragments. Instead, we found a preponderance of 3 MDa complexes (90 subunits) and no fragments smaller than 3 MDa. We also found 90‐mers in the disassembly of uncrosslinked HBV capsids. 90‐mers seem to be a common pause point in disassembly reactions. Partly explaining this result, graph theory simulations have showed a threshold for capsid stability between 80 and 90 subunits. To test a molecular breadboard concept, we showed that missing subunits could be refilled resulting in chimeric, 120 subunit particles. This result may be a means of assembling unique capsids with functional decorations. |
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| Keywords: | self‐assembly nanofluidics charge detection mass spectrometry resistive pulse sensing disassembly |
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