Analyzing macromolecular structures with native mass spectrometry: Insights to virus structure and assembly

ITQB- Seminar

Title: Analyzing macromolecular structures with native mass
spectrometry: Insights to virus structure and assembly

Speaker: Charlotte Uetrecht

Affiliation: Biomolecular Mass Spectrometry and Proteomics Group
Bijvoet Center for Biomolecular Research and Utrecht Institute for
Pharmaceutical Sciences
University of Utrecht

Host: Maria Arménia Carrondo- Structural Genomics

Abstract:

Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center and
UIPS, Utrecht University, The Netherlands and Netherlands Proteomics Centre
In recent years, native mass spectrometry has become a versatile tool in
structural biology. The invention of electrospray ionization enabled the
analysis of intact biomolecules and even noncovalent complexes by mass
spectrometry. Especially, the high sensitivity and broad mass range
facilitate the analysis of large protein complexes and their assembly
pathways.
Here, we studied the biophysical properties of Hepatitis B virus (HBV)
capsids. These viral protein shells are formed by multiple copies of a
single protein and exhibit icosahedral geometry. A remarkable feature of
the HBV capsid protein is the assembly into two distinct
stoichiometries. Using native mass spectrometry, the assembly of both
variants was shown to be complete whereas they differ in stability. The
capsids display a high stability in the gas phase retaining their
spherical structure as determined in ion mobility experiments.
Combination of this technique with computational modeling enabled the
identification of potential assembly intermediates. Furthermore,
dynamics like the exchange of subunits from the capsids with free
building blocks in solution can be monitored indicating the high
potential of mass spectrometric applications in structural biology and
biophysics not only in virology.