1.8 Å resolution structure of β-galactosidase with a 200 kV CRYO ARM electron microscope

IUCrJ
Alan MerkSriram Subramaniam

Abstract

We report the determination of the structure of Escherichia coli β-galactosidase at a resolution of ∼1.8 Å using data collected on a 200 kV CRYO ARM microscope equipped with a K3 direct electron detector. The data were collected in a single 24 h session by recording images from an array of 7 × 7 holes at each stage position using the automated data collection program SerialEM. In addition to the expected features such as holes in the densities of aromatic residues, the map also shows density bumps corresponding to the locations of hydrogen atoms. The hydrogen densities are useful in assigning absolute orientations for residues such as glutamine or asparagine by removing the uncertainty in the fitting of the amide groups, and are likely to be especially relevant in the context of structure-guided drug design. These findings validate the use of electron microscopes operating at 200 kV for imaging protein complexes at atomic resolution using cryo-EM.

References

Apr 6, 2006·Bioinformatics·Christian X Weichenberger, Manfred J Sippl
Jan 5, 2014·Journal of Structural Biology·Scott M StaggMichael S Chapman
Jul 30, 2014·Proceedings of the National Academy of Sciences of the United States of America·Alberto BartesaghiSriram Subramaniam
May 31, 2016·Cell·Alan MerkSriram Subramaniam
May 15, 2018·Structure·Alberto BartesaghiSriram Subramaniam
Sep 9, 2018·Nature Communications·Yong Zi TanDmitry Lyumkis
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May 13, 2019·Trends in Biochemical Sciences·Radostin DanevMasahide Kikkawa
Nov 12, 2019·IUCrJ·K NaydenovaC J Russo

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Citations

Dec 17, 2020·Frontiers in Cell and Developmental Biology·Minkyo JungJi Young Mun
Jul 3, 2021·International Journal of Molecular Sciences·Chun-Hsiung WangWei-Hau Chang
Aug 20, 2021·Proceedings of the National Academy of Sciences of the United States of America·Chancievan ThangaratnarajahDirk J Slotboom

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Methods Mentioned

BETA
size-exclusion chromatography
X-ray

Software Mentioned

SerialEM
CTFFIND
MotionCor2
RELION

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