The big chill: Growth of in situ structural biology with cryo-electron tomography DOI Creative Commons
Mikhail Kudryashev

QRB Discovery, Journal Year: 2024, Volume and Issue: 5

Published: Jan. 1, 2024

Abstract In situ structural biology with cryo-electron tomography (cryo-ET) and subtomogram averaging (StA) is evolving as a major method to understand the structure, function, interactions of biological molecules in cells single experiment. Since its inception, has matured some stellar highlights further opportunities broaden applications. this short review, I want provide personal perspective on developments cryo-ET have seen it for last ~20 years outline steps that led success. This my eyes junior researcher view present past hardware software cryo-ET.

Language: Английский

A protein blueprint of the diatom CO2-fixing organelle DOI Creative Commons
Onyou Nam,

Sabina Musiał,

Manon Demulder

et al.

Cell, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 1, 2024

Language: Английский

Citations

8

Diatom pyrenoids are encased in a protein shell that enables efficient CO2 fixation DOI Creative Commons
Ginga Shimakawa, Manon Demulder, Serena Flori

et al.

Cell, Journal Year: 2024, Volume and Issue: 187(21), P. 5919 - 5934.e19

Published: Oct. 1, 2024

Language: Английский

Citations

7

In-cell architecture of the mitochondrial respiratory chain DOI Creative Commons
Florent Waltz, Ricardo D. Righetto,

Ron Kelley

et al.

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

Published: Sept. 4, 2024

Abstract Mitochondria produce energy through oxidative phosphorylation, carried out by five membrane-bound complexes collectively known as the respiratory chain. These work in concert to transfer electrons and pump protons, leading ATP regeneration. The precise organization of these native cells is debated, notably their assembly into higher-order supercomplexes called respirasomes. Here, we use situ cryo-electron tomography visualize structures several major mitochondrial inside Chlamydomonas reinhardtii cells. synthases are segregated curved flat crista membrane domains, respectively. Respiratory I, III, IV assemble a single type respirasome, from which determined 5 Å-resolution structure showing binding electron carrier cytochrome c . Combined with single-particle microscopy reconstruction at 2.4 Å resolution, detailed model how interact each other mitochondria.

Language: Английский

Citations

2

A Protein Blueprint of the Diatom CO2-Fixing Organelle DOI Creative Commons
Onyou Nam, Sabina Musiał, Manon Demulder

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 26, 2023

Abstract Diatoms are central to the global carbon cycle. At heart of diatom fixation is an overlooked organelle called pyrenoid, where concentrated CO 2 delivered densely packed Rubisco. Diatom pyrenoids fix approximately one-fifth , but virtually nothing known about this in diatoms. Using large-scale fluorescence protein tagging and affinity purification-mass spectrometry, we generate a high-confidence spatially-defined protein-protein interaction network for pyrenoid. Within our pyrenoid 10 proteins with no function. We show that six these form static shell encapsulating Rubisco matrix critical structural integrity, shape, Although not conserved at sequence level, shares some architectural similarities prokaryotic carboxysomes. Collectively, results support convergent evolution across two main plastid lineages uncover major functional component fixation.

Language: Английский

Citations

4

Molecular architecture of thylakoid membranes within intact spinach chloroplasts DOI Creative Commons
Wojciech Wietrzyñski, Lorenz Lamm, William H. Wood

et al.

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

Published: Nov. 26, 2024

Abstract Thylakoid membranes coordinate the light reactions of photosynthesis across multiple scales, coupling architecture an elaborate membrane network to spatial organization individual protein complexes embedded within this network. Previously, we used in situ cryo- electron tomography (cryo-ET) reveal native thylakoid green alga Chlamydomonas reinhardtii [1] and then map molecular these thylakoids with single-molecule precision [2]. However, it remains be shown how generalizable algal blueprint is vascular plants, which possess distinct subdivided into grana stacks interconnected by non-stacked stromal lamellae. Here, continue our cryo-ET investigation intact chloroplasts isolated from spinach ( Spinacia oleracea ). We visualize fine ultrastructural details membranes, as well interactions between plastoglobules. apply further develop AI-based computational approaches for automated segmentation picking [3], enabling us quantify photosynthetic plane adjacent stacked membranes. Our analysis reveals that, despite different 3D architecture, plants algae strikingly similar. In contrast plant thylakoids, where semi- crystalline arrays photosystem II (PSII) appear hold some together, find that PSII non-crystalline has uniform concentration both Similar C. , observe strict lateral heterogeneity PSI at boundary appressed non-appressed domains, no evidence a margin region have been proposed intermix. Based on measurements, support simple two-domain model plants.

Language: Английский

Citations

0

The big chill: Growth of in situ structural biology with cryo-electron tomography DOI Creative Commons
Mikhail Kudryashev

QRB Discovery, Journal Year: 2024, Volume and Issue: 5

Published: Jan. 1, 2024

Abstract In situ structural biology with cryo-electron tomography (cryo-ET) and subtomogram averaging (StA) is evolving as a major method to understand the structure, function, interactions of biological molecules in cells single experiment. Since its inception, has matured some stellar highlights further opportunities broaden applications. this short review, I want provide personal perspective on developments cryo-ET have seen it for last ~20 years outline steps that led success. This my eyes junior researcher view present past hardware software cryo-ET.

Language: Английский

Citations

0