Erratum to: Crystallization in Microgravity and the Atomic-Resolution Structure of Uridine Phosphorylase from Vibrio cholerae

AbstractThe pathway for the biosynthesis of the bacterial cell wall is one of the most prolific antibiotic targets, exemplified by the widespread use of β-lactam antibiotics. Despite this, our structural understanding of class A penicillin binding proteins, which perform the last two steps in this pathway, is incomplete due to the inherent difficulty in their crystallization and the complexity of their substrates. Here, we determine the near atomic resolution structure of the 83 kDa class A PBP from Escherichia coli, PBP1b, using cryogenic electron microscopy and a styrene maleic acid anhydride membrane mimetic. PBP1b, in its apo form, is seen to exhibit a distinct conformation in comparison to Moenomycin-bound crystal structures. The work herein paves the way for the use of cryoEM in structure-guided antibiotic development for this notoriously difficult to crystalize class of proteins and their complex substrates.

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A resolution record for cryoEM

Faculty Reviews ◽

10.12703/r-01-000002 ◽

2021 ◽

Vol 10 ◽

Author(s):

Jonathan Ashmore ◽

Bridget Carragher ◽

Peter B Rosenthal ◽

William Weis

Keyword(s):

Electron Microscopy ◽

Image Analysis ◽

Structure Determination ◽

Test Specimen ◽

Structural Information ◽

Atomic Resolution ◽

Fast Growing ◽

Cryo Electron Microscopy ◽

New Instrument ◽

Resolution Structure

Cryo electron microscopy (cryoEM) is a fast-growing technique for structure determination. Two recent papers report the first atomic resolution structure of a protein obtained by averaging images of frozen-hydrated biomolecules. They both describe maps of symmetric apoferritin assemblies, a common test specimen, in unprecedented detail. New instrument improvements, different in the two studies, have contributed better images, and image analysis can extract structural information sufficient to resolve individual atomic positions. While true atomic resolution maps will not be routine for most proteins, the studies suggest structures determined by cryoEM will continue to improve, increasing their impact on biology and medicine.

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A Covalent Modification of NADP+Revealed by the Atomic Resolution Structure of FprA, aMycobacterium tuberculosisOxidoreductase†,‡

Biochemistry ◽

10.1021/bi025858a ◽

2002 ◽

Vol 41 (28) ◽

pp. 8807-8818 ◽

Cited By ~ 31

Author(s):

Roberto T. Bossi ◽

Alessandro Aliverti ◽

Debora Raimondi ◽

Federico Fischer ◽

Giuliana Zanetti ◽

...

Keyword(s):

Covalent Modification ◽

Atomic Resolution ◽

Resolution Structure

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Atomic-resolution structure of cytoskeletal bactofilin by solid-state NMR

Science Advances ◽

10.1126/sciadv.1501087 ◽

2015 ◽

Vol 1 (11) ◽

pp. e1501087 ◽

Cited By ~ 39

Author(s):

Chaowei Shi ◽

Pascal Fricke ◽

Lin Lin ◽

Veniamin Chevelkov ◽

Melanie Wegstroth ◽

...

Keyword(s):

Solid State ◽

Solid State Nmr ◽

Caulobacter Crescentus ◽

Heavy Atom ◽

Cytoskeletal Proteins ◽

Atomic Resolution ◽

Hydrophobic Core ◽

X Ray Crystallography ◽

Wide Range ◽

Resolution Structure

Bactofilins are a recently discovered class of cytoskeletal proteins of which no atomic-resolution structure has been reported thus far. The bacterial cytoskeleton plays an essential role in a wide range of processes, including morphogenesis, cell division, and motility. Among the cytoskeletal proteins, the bactofilins are bacteria-specific and do not have a eukaryotic counterpart. The bactofilin BacA of the speciesCaulobacter crescentusis not amenable to study by x-ray crystallography or solution nuclear magnetic resonance (NMR) because of its inherent noncrystallinity and insolubility. We present the atomic structure of BacA calculated from solid-state NMR–derived distance restraints. We show that the core domain of BacA forms a right-handed β helix with six windings and a triangular hydrophobic core. The BacA structure was determined to 1.0 Å precision (heavy-atom root mean square deviation) on the basis of unambiguous restraints derived from four-dimensional (4D) HN-HN and 2D C-C NMR spectra.

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