In situstructural analysis of mammalian cells using a 200 kV electron cryomicroscope – implications for research infrastructure DOI Creative Commons
Piotr Szwedziak

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 11, 2024

Abstract Electron cryotomography is a powerful imaging technique allowing for studying functional cellular modules in their native environment with macromolecular resolution. However, it requires access to complex and expensive instrumentation, typically 300 kV electron cryomicroscope equipped an energy filter. Simpler cheaper 200 100 instruments have been successfully used single particle cryoEM analyses, which has helped democratize the broaden access. Here, we set out investigate if without filter can be utilized situ structural studies of mammalian cells by cryotomography. We demonstrate that resulting tomograms thin edges U2OS are sufficient quality annotate contents cell observe spatial inter-relationships among macromolecules. In particular, undertook molecular sociology analysis lipid droplets addressed subcellular distribution interactions other organelles. Finally, examined geographical scientific output two most common cryomicroscopy platforms deduced heavily underutilized respect applications. Given favorable ratio versatility versus costs foresee cryomicroscopes will become workhorses local facilities.

Language: Английский

Protein nanoparticle vaccines induce potent neutralizing antibody responses against MERS-CoV DOI Creative Commons
Cara W. Chao, Kaitlin R. Sprouse, Marcos C. Miranda

et al.

Cell Reports, Journal Year: 2024, Volume and Issue: 43(12), P. 115036 - 115036

Published: Dec. 1, 2024

Language: Английский

Citations

4
Structure-guided design of a prefusion GPC trimer induces neutralizing responses against LASV DOI Creative Commons

Shaoyan Wang,

Ruihua Li, Xiaoyan Pan

et al.

npj Vaccines, Journal Year: 2025, Volume and Issue: 10(1)

Published: Feb. 22, 2025

Lassa virus (LASV) belongs to the Arenaviridae family and causes severe hemorrhagic fever in humans. Although many vaccine candidates for exist, no vaccines have been approved clinical use currently. The precursor glycoprotein complex (GPC), which is expressed as a trimer on viral surface, main target development. However, it has significant challenge elicit effective neutralizing antibodies against LASV. In this study, we designed produced prefusion GPC antigen of LASV, named GPCv2. Based structural information GPC, made modifications by replacing amino acid at position 328 with proline appending trimerization domain. This resulted highly trimeric form GPCv2 that retained important conformational epitopes stimulated higher levels antibodies. Moreover, vaccination protected mice from LASV pseudovirus challenge. Additionally, immune repertoire sequencing showed induced clones group were more convergent its own unique V-J pairing bias compared monomeric group. These findings demonstrate potential promising candidate an

Language: Английский

Citations

0
In situstructural analysis of mammalian cells using a 200 kV electron cryomicroscope – implications for research infrastructure DOI Creative Commons
Piotr Szwedziak

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 11, 2024

Abstract Electron cryotomography is a powerful imaging technique allowing for studying functional cellular modules in their native environment with macromolecular resolution. However, it requires access to complex and expensive instrumentation, typically 300 kV electron cryomicroscope equipped an energy filter. Simpler cheaper 200 100 instruments have been successfully used single particle cryoEM analyses, which has helped democratize the broaden access. Here, we set out investigate if without filter can be utilized situ structural studies of mammalian cells by cryotomography. We demonstrate that resulting tomograms thin edges U2OS are sufficient quality annotate contents cell observe spatial inter-relationships among macromolecules. In particular, undertook molecular sociology analysis lipid droplets addressed subcellular distribution interactions other organelles. Finally, examined geographical scientific output two most common cryomicroscopy platforms deduced heavily underutilized respect applications. Given favorable ratio versatility versus costs foresee cryomicroscopes will become workhorses local facilities.

Language: Английский

Citations

0