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2014 ; 346
(6215
): 1377-80
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Electron microscopy: Ultrastable gold substrates for electron cryomicroscopy
#MMPMID25504723
Russo CJ
; Passmore LA
Science
2014[Dec]; 346
(6215
): 1377-80
PMID25504723
show ga
Despite recent advances, the structures of many proteins cannot be determined by
electron cryomicroscopy because the individual proteins move during irradiation.
This blurs the images so that they cannot be aligned with each other to calculate
a three-dimensional density. Much of this movement stems from instabilities in
the carbon substrates used to support frozen samples in the microscope. Here we
demonstrate a gold specimen support that nearly eliminates substrate motion
during irradiation. This increases the subnanometer image contrast such that ?
helices of individual proteins are resolved. With this improvement, we determine
the structure of apoferritin, a smooth octahedral shell of ?-helical subunits
that is particularly difficult to solve by electron microscopy. This advance in
substrate design will enable the solution of currently intractable protein
structures.