Biology subjects

von Kuegelgen, A.

Publications and source records attributed to von Kuegelgen, A..

4 recordsLinked to original sources

bioRxiv2026

Molecular architecture of colossal surface layers from hyperthermophilic archaea

Surface layers (S-layers) are paracrystalline protein lattices that form the outermost layer of the cell envelope in most archaea, providing structural support, protecting against external insults, and co-ordinating interactions with their environment. Despite their widespread occurrence, the molecular and structural details of S-layer architecture in hyperthermophilic archaea remain largely unknown. Here, we report the structure and cellular architecture of the S-layer from the hyperthermophilic archaeon Pyrobaculum arsenaticum by combining in situ electron cryotomography with single-particle electron cryomicroscopy, AlphaFold modelling, and peptide-fingerprinting mass spectrometry. We show that the S-layer is formed by an uncharacterised 292-kDa S-layer protein (SLP) extending 37 nm from the cytoplasmic membrane, making it, to our knowledge, the largest SLP structurally characterised to date. This SLP has a remarkable multidomain architecture comprising 19 immunoglobulin-like domains, 14 canonical and five non-canonical, organised into a lattice-forming core, a stalk, and a unique crown domain that stabilise the S-layer. Comparative genomic analyses unearthed homologous colossal SLP candidates across Thermoproteota, indicating that this distinctive architecture is conserved across diverse archaeal lineages. Together, our findings provide a structural framework for understanding the cell-surface organisation in hyperthermophilic archaea and suggest that these colossal S-layers represent a specialised adaptation to life at high temperatures.

microbiology↗

bioRxiv2025

An Asgard archaeon with internal membrane compartments

The emergence of eukaryotes from a merger between an archaeon and a bacterial cell [~]two billion years ago involved a profound change in cellular organisation. While the order in which different features of the eukaryotic cell arose remains a matter of controversy1-3, close archaeal relatives of eukaryotes have recently been identified that possess homologues of eukaryotic trafficking machinery4,5 and a complex cell architecture6,7. The members of this phylum, the Asgard archaea (syn. Prometheoarchaeota) described so far, however, lack internal membrane-bound compartments, and therefore have shed little light on origins of the hallmark eukaryotic endomembrane system. Here we report the cell biological analysis of a member of the Heimdallarchaeia, Candidatus Y. umbracryptum in enriched mixed microbial communities. Possessing a small genome encoding few homologues of eukaryotic membrane remodelling machinery8, Ca. Y. umbracryptum cells in late-stage cultures resemble previously described Asgard archaea with extensive cellular protrusions. Surprisingly, during early stages of culture growth Ca. Y. umbracryptum cells have fewer protrusions but possess numerous intracellular vesicles, most of which have a luminal surface that morphologically resembles the outer coat of the plasma membrane. These data alter our view of eukaryogenesis by identifying a close archaeal relative of eukaryotes with an endomembrane system, whose appearance changes with cell state.

cell biology↗

bioRxiv2022

Interdigitated immunoglobulin arrays form the hyperstable surface layer of the extremophilic bacterium Deinococcus radiodurans

Deinococcus radiodurans is an atypical diderm bacterium with a remarkable ability to tolerate various environmental stresses, partly because of its complex cell envelope encapsulated within a hyperstable surface layer (S-layer). Despite decades of research into this cell envelope, atomic structural details of the S-layer have remained obscure. In this study, we report the electron cryomicroscopy structure of the D. radiodurans S-layer, showing how it is formed by the Hexagonally Packed Intermediate-layer (HPI) protein arranged in a planar hexagonal lattice. The HPI protein forms an array of immunoglobulin-like folds within the S-layer, with each monomer extending into the adjoining hexamer, leading to a highly interconnected, stable, sheet-like arrangement. Using electron cryotomography and subtomogram averaging from focused ion beam-milled D. radiodurans cells, we obtained a structure of the cellular S-layer, showing how this HPI S-layer coats native membranes on the surface of cells. Our S-layer structure from the diderm bacterium D. radiodurans shows similarities to immunoglobulin-like domain-containing S-layers from monoderm bacteria and archaea, highlighting shared traits in cell surface organization across different domains of life, with connotations on the evolution of immunoglobulin-based molecular recognition systems in eukaryotes.

microbiology↗

bioRxiv2022

A multi-domain connector links the outer membrane and cell wall in deep-branching bacteria

Deinococcus radiodurans is a deep-branching extremophilic bacterium that is remarkably tolerant to numerous environmental stresses, including large doses of ultraviolet radiation and extreme temperatures. It can even survive in outer space for several years. This endurance of D. radiodurans has been partly ascribed to its atypical cell envelope comprising an inner membrane, a large periplasmic space with a thick peptidoglycan (PG) layer, and an outer membrane (OM) covered by a surface layer (S-layer). Despite intense research, molecular principles governing envelope organization and OM stabilization are unclear in D. radiodurans and related bacteria. Here, we report an electron cryomicroscopy (cryo-EM) structure of the abundant D. radiodurans OM protein SlpA, showing how its C-terminal segment forms homotrimers of 30-stranded {beta}-barrels in the OM, whereas its N-terminal segment forms long, homotrimeric coiled coils linking the OM to the PG layer via S-layer homology (SLH) domains. Using the power of structure prediction and sequence-based bioinformatics, we further show that SlpA-like proteins are widespread in deep-branching Gram-negative bacteria, plausibly constituting an ancestral superfamily of OM-PG connectors, important for organizing the cell envelopes of many bacteria. Finally, combining our atomic structures with tomography of cell envelopes, we report a model for the cell surface of D. radiodurans, with implications on understanding the cell surface organization and hyperstability of D. radiodurans and related bacteria. Furthermore, the widespread occurrence of SlpA-like OM-PG connectors in deep-branching bacteria will help in understanding the evolutionary transition between Gram-negative and Gram-positive bacteria.

microbiology↗