Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies

Klein, Steffen; Wimmer, Benedikt; Winter, Sophie L.; Kolovou, Androniki; Laketa, Vibor; Chlanda, Petr

doi:10.1038/s42003-020-01567-z

Cited by 43 publications

(27 citation statements)

References 51 publications

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“…Due to the disruption of membranes in lysed cells, the topology of these complexes is difficult to unravel. However, these structures share a morphological similarity to a membrane-associated dome protein complex recently described on the limiting membrane of the lamellar bodies inside alveolar cells [44].…”

Section: Discussionsupporting

confidence: 56%

In situ imaging of bacterial outer membrane projections and associated protein complexes using electron cryo-tomography

Kaplan

Chreifi

Metskas

et al. 2021

eLife

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The ability to produce outer membrane projections in the form of tubular membrane extensions (MEs) and membrane vesicles (MVs) is a widespread phenomenon among diderm bacteria. Despite this, our knowledge of the ultrastructure of these extensions and their associated protein complexes remains limited. Here, we surveyed the ultrastructure and formation of MEs and MVs, and their associated protein complexes, in tens of thousands of electron cryo-tomograms of ~ 90 bacterial species that we have collected for various projects over the past 15 years (Jensen lab database), in addition to data generated in the Briegel lab. We identified outer MEs and MVs in 13 diderm bacterial species and classified several major ultrastructures: 1) tubes with a uniform diameter (with or without an internal scaffold), 2) tubes with irregular diameter, 3) tubes with a vesicular dilation at their tip, 4) pearling tubes, 5) connected chains of vesicles (with or without neck-like connectors), 6) budding vesicles and nanopods. We also identified several protein complexes associated with these MEs and MVs which were distributed either randomly or exclusively at the tip. These complexes include a secretin-like structure and a novel crown-shaped structure observed primarily in vesicles from lysed cells. In total, this work helps to characterize the diversity of bacterial membrane projections and lays the groundwork for future research in this field.

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Section: Discussionsupporting

confidence: 56%

In situ imaging of bacterial outer membrane projections and associated protein complexes using electron cryo-tomography

Kaplan

Chreifi

Metskas

et al. 2021

eLife

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show abstract

Section: Discussionsupporting

confidence: 56%

In situ imaging of bacterial membrane projections and associated protein complexes using electron cryo-tomography

Kaplan

Chreifi

Metskas

et al. 2021

Preprint

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The ability to produce membrane projections in the form of tubular membrane extensions (MEs) and membrane vesicles (MVs) is a widespread phenomenon among bacteria. Despite this, our knowledge of the ultrastructure of these extensions and their associated protein complexes remains limited. Here, we surveyed the ultrastructure and formation of MEs and MVs, and their associated protein complexes, in tens of thousands of electron cryo-tomograms of ~ 90 bacterial species that we have collected for various projects over the past 15 years (Jensen lab database), in addition to data generated in the Briegel lab. We identified MEs and MVs in 13 species and classified several major ultrastructures: 1) tubes with a uniform diameter (with or without an internal scaffold), 2) tubes with irregular diameter, 3) tubes with a vesicular dilation at their tip, 4) pearling tubes, 5) connected chains of vesicles (with or without neck-like connectors), 6) budding vesicles and nanopods. We also identified several protein complexes associated with these MEs and MVs which were distributed either randomly or exclusively at the tip. These complexes include a secretin-like structure and a novel crown-shaped structure observed primarily in vesicles from lysed cells. In total, this work helps to characterize the diversity of bacterial membrane projections and lays the groundwork for future research in this field.

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“…In a third approach, Klein et al. ( Klein et al., 2021 ) collected cryo-fluorescence data both before milling, to target cells for lamella preparation, and again after cryo-ET data collection. By this method, the fluorescence data collected from the milled lamella could be superimposed back on the tomographic reconstruction.…”

Section: Identifying Structures and Targeted Millingmentioning

confidence: 99%

Challenges and triumphs in cryo-electron tomography

Hylton

Swulius

2021

iScience

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Summary Cryo-electron tomography has stepped fully into the spotlight. Enthusiasm is high. Fortunately for us, this is an exciting time to be a cryotomographer, but there is still a way to go before declaring victory. Despite its potential, cryo-electron tomography possesses many inherent challenges. How do we image through thick cell samples, and possibly even tissue? How do we identify a protein of interest amidst the noisy, crowded environment of the cytoplasm? How do we target specific moments of a dynamic cellular process for tomographic imaging? In this review, we cover the history of cryo-electron tomography and how it came to be, roughly speaking, as well as the many approaches that have been developed to overcome its intrinsic limitations.

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Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies

Cited by 43 publications

References 51 publications

In situ imaging of bacterial outer membrane projections and associated protein complexes using electron cryo-tomography

In situ imaging of bacterial outer membrane projections and associated protein complexes using electron cryo-tomography

In situ imaging of bacterial membrane projections and associated protein complexes using electron cryo-tomography

Challenges and triumphs in cryo-electron tomography

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