@article {69687,
	title = {Structural dynamics of human ribosomes in situ reconstructed by exhaustive high-resolution template matching.},
	journal = {Mol Cell},
	year = {2024},
	month = {2024 Nov 25},
	abstract = {<p>Protein synthesis is central to life and requires the ribosome, which catalyzes the stepwise addition of amino acids to a polypeptide chain by undergoing a sequence of structural transformations. Here, we employed high-resolution template matching (HRTM) on cryoelectron microscopy (cryo-EM) images of directly cryofixed living cells to obtain a set of ribosomal configurations covering the entire elongation cycle, with each configuration occurring at its native abundance. HRTM{\textquoteright}s position and orientation precision and ability to detect small targets (\~{}300\&nbsp;kDa) made it possible to order these configurations along the reaction coordinate and to reconstruct molecular features of any configuration along the elongation cycle. Visualizing the cycle{\textquoteright}s structural dynamics by combining a sequence of \&gt;40 reconstructions into a 3D movie readily revealed component and ligand movements, some of them surprising, such as spring-like intramolecular motion, providing clues about the molecular mechanisms involved in some still mysterious steps during chain elongation.</p>
},
	issn = {1097-4164},
	doi = {10.1016/j.molcel.2024.11.003},
	author = {Rickgauer, J Peter and Choi, Heejun and Moore, Andrew S and Denk, Winfried and Lippincott-Schwartz, Jennifer}
}
@article {48460,
	title = {Single-protein detection in crowded molecular environments in cryo-EM images.},
	journal = {eLife},
	volume = {6},
	year = {2017},
	month = {2017 May 03},
	abstract = {<p>We present an approach to study macromolecular assemblies by detecting component proteins{\textquoteright} characteristic high-resolution projection patterns, calculated from their known 3D structures, in single electron cryo-micrographs. Our method detects single apoferritin molecules in vitreous ice with high specificity and determines their orientation and location precisely. Simulations show that high spatial-frequency information and-in the presence of protein background-a whitening filter are essential for optimal detection, in particular for images taken far from focus. Experimentally, we could detect small viral RNA polymerase molecules, distributed randomly among binding locations, inside rotavirus particles. Based on the currently attainable image quality, we estimate a threshold for detection that is 150 kDa in ice and 300 kDa in 100 nm thick samples of dense biological material.</p>
},
	issn = {2050-084X},
	doi = {10.7554/eLife.25648},
	author = {Rickgauer, J Peter and Grigorieff, Nikolaus and Denk, Winfried}
}
