TY - JOUR
T1 - 2.2 å resolution cryo-EM structure of β-galactosidase in complex with a cell-permeant inhibitor
AU - Bartesaghi, Alberto
AU - Merk, Alan
AU - Banerjee, Soojay
AU - Matthies, Doreen
AU - Wu, Xiongwu
AU - Milne, Jacqueline L S
AU - Subramaniam, Sriram
PY - 2015/6/5
Y1 - 2015/6/5
N2 - Cryo-electron microscopy (cryo-EM) is rapidly emerging as a powerful tool for protein structure determination at high resolution. Here we report the structure of a complex between Escherichia coli b-galactosidase and the cell-permeant inhibitor phenylethyl b-D-thiogalactopyranoside (PETG), determined by cryo-EM at an average resolution of ∼2.2 angstroms (å). Besides the PETG ligand, we identified densities in the map for ∼800 water molecules and for magnesium and sodium ions. Although it is likely that continued advances in detector technology may further enhance resolution, our findings demonstrate that preparation of specimens of adequate quality and intrinsic protein flexibility, rather than imaging or image-processing technologies, now represent the major bottlenecks to routinely achieving resolutions close to 2 å using single-particle cryo-EM.
AB - Cryo-electron microscopy (cryo-EM) is rapidly emerging as a powerful tool for protein structure determination at high resolution. Here we report the structure of a complex between Escherichia coli b-galactosidase and the cell-permeant inhibitor phenylethyl b-D-thiogalactopyranoside (PETG), determined by cryo-EM at an average resolution of ∼2.2 angstroms (å). Besides the PETG ligand, we identified densities in the map for ∼800 water molecules and for magnesium and sodium ions. Although it is likely that continued advances in detector technology may further enhance resolution, our findings demonstrate that preparation of specimens of adequate quality and intrinsic protein flexibility, rather than imaging or image-processing technologies, now represent the major bottlenecks to routinely achieving resolutions close to 2 å using single-particle cryo-EM.
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U2 - 10.1126/science.aab1576
DO - 10.1126/science.aab1576
M3 - Article
C2 - 25953817
AN - SCOPUS:84930667920
SN - 0036-8075
VL - 348
SP - 1147
EP - 1151
JO - Science
JF - Science
IS - 6239
ER -