Dataset Information

Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED.

ABSTRACT: Traditionally, crystallographic analysis of macromolecules has depended on large, well-ordered crystals, which often require significant effort to obtain. Even sizable crystals sometimes suffer from pathologies that render them inappropriate for high-resolution structure determination. Here we show that fragmentation of large, imperfect crystals into microcrystals or nanocrystals can provide a simple path for high-resolution structure determination by the cryoEM method MicroED and potentially by serial femtosecond crystallography.

SUBMITTER: de la Cruz MJ

PROVIDER: S-EPMC5376236 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED.

de la Cruz M Jason MJ Hattne Johan J Shi Dan D Seidler Paul P Rodriguez Jose J Reyes Francis E FE Sawaya Michael R MR Cascio Duilio D Weiss Simon C SC Kim Sun Kyung SK Hinck Cynthia S CS Hinck Andrew P AP Calero Guillermo G Eisenberg David D Gonen Tamir T

Nature methods 20170201 4

Traditionally, crystallographic analysis of macromolecules has depended on large, well-ordered crystals, which often require significant effort to obtain. Even sizable crystals sometimes suffer from pathologies that render them inappropriate for high-resolution structure determination. Here we show that fragmentation of large, imperfect crystals into microcrystals or nanocrystals can provide a simple path for high-resolution structure determination by the cryoEM method MicroED and potentially by ...[more]

PMID: 28192420

Similar Datasets

Project description:Structural studies of proteins usually rely on a model obtained from one crystal. By investigating the details of this model, crystallographers seek to obtain insight into the function of the macromolecule. It is therefore important to know which details of a protein structure are reproducible or to what extent they might differ. To address this question, the high-resolution structures of five crystals of bovine trypsin obtained under analogous conditions were compared. Global parameters and structural details were investigated. All of the models were of similar quality and the pairwise merged intensities had large correlation coefficients. The C(?) and backbone atoms of the structures superposed very well. The occupancy of ligands in regions of low thermal motion was reproducible, whereas solvent molecules containing heavier atoms (such as sulfur) or those located on the surface could differ significantly. The coordination lengths of the calcium ion were conserved. A large proportion of the multiple conformations refined to similar occupancies and the residues adopted similar orientations. More than three quarters of the water-molecule sites were conserved within 0.5?Å and more than one third were conserved within 0.1?Å. An investigation of the protonation states of histidine residues and carboxylate moieties was consistent for all of the models. Radiation-damage effects to disulfide bridges were observed for the same residues and to similar extents. Main-chain bond lengths and angles averaged to similar values and were in agreement with the Engh and Huber targets. Other features, such as peptide flips and the double conformation of the inhibitor molecule, were also reproducible in all of the trypsin structures. Therefore, many details are similar in models obtained from different crystals. However, several features of residues or ligands located in flexible parts of the macromolecule may vary significantly, such as side-chain orientations and the occupancies of certain fragments.

Dataset Information

Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED.

Publications

Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets