Research Projects
Protein Dynamics
The solution-phase protein motion that is part of a multi-component complex can not always be inferred from the three-dimensional structure. For example, in contrast to the still-life representation of viral capsids in models based on cryo-electron microscopy and X-ray crystallography, these supramolecular protein complexes are highly dynamic in solution. The range and frequency of capsid protein dynamics are poorly understood, despite evidence that the infectivity of animal viruses requires conformational freedom. Protein function is intimately connected to dynamics and therefore knowledge of the frequency, range, and coordination of motion by supramolecular complexes is critical to understanding how they function. Our lab uses viruses as a paradigm for studying protein dynamics in supramolecular complexes. With the use of kinetic hydrolysis and quantitative mass spectrometry, we are determining the free energy and rates of large scale protein motion within viral particles. These are the first quantitative measurements for protein dynamics in a megadalton complex. Hydrogen-deuterium exchange, chemical labeling, and quartz crystal microbalance measurements are a few of the additional methods applied to the quantitative analysis of virus particle stability and dynamics. Graduate students: Jonathan Hilmer and Vamseedhar Rayaprolu. Undergraduates: Lena, Petersen and Tim Potter. Technical assistance: Geoff Blatter
Publications
Bothner B, Taylor DJ, Jun B, Lee KK, Siuzdak G, Schultz CP, Johnson JE
, "Maturation of a tetravirus capsid alters the dynamic properties and creates a metastable complex." Virology 334(1):17-27 (2005)
Lee KK, Tang J, Taylor D, Bothner B, Johnson JE
, "Small Compounds Targeted to Subunit Interfaces Arrest Maturation in a Nonenveloped, Icosahedral Animal Virus." J. Virology 78(13): 7208-7216 (2004)
Taylor DJ, Wang Q, Bothner B, Natarajan P, Finn MG, Johnson JE, "Correlation of chemical reactivity of Nudaurelia capensis omega virus with a pH-induced conformational change." Chem Commun (Camb). 22: 2770-2771 (2003)
Bothner B, Schneemann A, Marshall D, Reddy V, Johnson JE, Siuzdak G, "Crystallographically identical virus capsids display different properties in solution." Nature Struct. Biol. 2:114-116 (1999)
Lewis JK, Bothner B, Smith TJ, Siuzdak G, "Antiviral agent blocks breathing of the common cold virus." Proc Natl Acad Sci U S A 95(12):6774-6778 (1998)
Bothner B, Dong XF, Bibbs L, Johnson JE, Siuzdak G, "Evidence of viral capsid dynamics using limited proteolysis and mass spectrometry." J Biol Chem 273(2):673-676 (1998)
Speir JA, Bothner B, Qu C, Willits DA, Young MJ, Johnson JE, "Enhanced Local Symmetry Interactions Globally Stabilize a Mutant Virus Capsid that Maintains Infectivity and Capsid Dynamics." J. Virology 80(7):3582-3591. (2006)
Hilmer JK, Zlotnick A, Bothner B., "Conformational equilibria and rates of
localized motion within hepatitis B virus capsids." J Mol Biol. 2008 Jan11;375(2):581-94. Epub 2007 Oct 22. PubMed PMID: 18022640; PubMed Central PMCID: PMC2238684. (2008)
Personnel:
Brian Bothner
|
