Polyelectrolyte coating of cryo-EM grids improves lateral distribution and prevents aggregation of macromolecules

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F22%3A00128514" target="_blank" >RIV/00216224:14740/22:00128514 - isvavai.cz</a>

Alternative codes found

RIV/62156489:43210/22:43922198

Result on the web

<a href="https://journals.iucr.org/d/issues/2022/11/00/jb5048/index.html" target="_blank" >https://journals.iucr.org/d/issues/2022/11/00/jb5048/index.html</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1107/S2059798322009299" target="_blank" >10.1107/S2059798322009299</a>

Result language

angličtina

Original language name

Polyelectrolyte coating of cryo-EM grids improves lateral distribution and prevents aggregation of macromolecules

Original language description

Cryo-electron microscopy (cryo-EM) is one of the primary methods used to determine the structures of macromolecules and their complexes. With the increased availability of cryo-electron microscopes, the preparation of highquality samples has become a bottleneck in the cryo-EM structure-determination pipeline. Macromolecules can be damaged during the purification or preparation of vitrified samples for cryo-EM, making them prone to binding to the grid support, to aggregation or to the adoption of preferential orientations at the airwater interface. Here, it is shown that coating cryo-EM grids with a negatively charged polyelectrolyte, such as single-stranded DNA, before applying the sample reduces the aggregation of macromolecules and improves their distribution. The single-stranded DNA-coated grids enabled the determination of high-resolution structures from samples that aggregated on conventional grids. The polyelectrolyte coating reduces the diffusion of macromolecules and thus may limit the negative effects of the contact of macromolecules with the grid support and blotting paper, as well as of the shear forces on macromolecules during grid blotting. Coating grids with polyelectrolytes can readily be employed in any laboratory dealing with cryo-EM sample preparation, since it is fast, simple, inexpensive and does not require specialized equipment.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2022

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Acta Crystallographica Section D: Structural Biology

ISSN

2059-7983

e-ISSN

—

Volume of the periodical

78

Issue of the periodical within the volume

NOV

Country of publishing house

GB - UNITED KINGDOM

Number of pages

10

Pages from-to

1337-1346

UT code for WoS article

000885880300005

EID of the result in the Scopus database

2-s2.0-85141889420