Ellipsoid localisation microscopy infers the size and order of protein layers in Bacillus spore coats
Ellipsoid Localisation Microscopy
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Manetsberger, J., Manton, J., Erdelyi, M., Lin, H., Rees, H., Christie, G., & Rees, E. (2015). Ellipsoid localisation microscopy infers the size and order of protein layers in Bacillus spore coats. Biophysical Journal, 109 2058-2066. https://doi.org/10.1016/j.bpj.2015.09.023
Multi-layered protein coats are crucial to the dormancy, robustness, and germination of bacterial spores. In Bacillus subtilis spores the coat contains over 70 distinct proteins. Identifying which proteins reside in each layer may provide insight into their distinct functions. We present image analysis methods which determine the order and geometry of concentric protein layers by fitting a model description for a spheroidal fluorescent shell image to optical micrographs of spores incorporating fluorescent fusion proteins. The radius of a spherical protein shell can be determined more precisely than 10 nm by fitting an equation to widefield fluorescence micrographs. Ellipsoidal shell axes can be fitted with comparable precision. The layer orders inferred for B. subtilis and B. megaterium are consistent with measurements in literature. The aspect ratio of elongated spores and the tendency of some proteins to localise near their poles can be quantified, enabling measurement of structural anisotropy.
We gratefully acknowledge support from MedImmune through the Beacon collaboration, the EPSRC Centre for Doctoral Training in Sensor Technologies and Applications (EP/L015889/1), and thank Clemens Kaminski, Romain Laine and Jose Casas-Finet for inspiring discussions.
External DOI: https://doi.org/10.1016/j.bpj.2015.09.023
This record's URL: https://www.repository.cam.ac.uk/handle/1810/251107
Creative Commons Attribution 4.0 International License
Licence URL: http://creativecommons.org/licenses/by/4.0/
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