Cell-surface protein glycosylation is a fundamental post-translational modification that plays a crucial role in membrane protein function and cellular behavior. However, elucidating the molecular spa...

Over recent decades, the glycocalyx, an extracellular organelle composed of a multitude of glycolipids, glycoproteins, proteoglycans, and glycoRNA, has gained considerable interest in cellular biology. While research in this field has revealed its tremendous importance in ever more aspects of physiological and pathological cellular processes, many of the principles that govern the role of the glycocalyx in these processes on a molecular level are still unknown. In order to unravel the fundamental laws underlying glycocalyx function, new technologies are required that enable the distinction between individual subprocesses within the intricate environment of the glycocalyx. Here, we establish an experimental platform to investigate the dynamics of the glycocalyx at the nanometer and microsecond length and time scales in a bottom-up fashion. We synthesized defined oligosaccharides and installed them on supported lipid bilayers. This way, synthetic glycolipids were assembled to glycocalyx model systems with tunable properties. By investigating these tunable model systems with interferometric scattering (iSCAT) microscopy, we gain access to the required spatiotemporal resolution. We found a strong correlation between the molecular structure of several investigated model glycans and global dynamics of the system. Our findings are corroborated by atomistic molecular dynamics simulations and coarse-grained Brownian dynamics simulations. Our results provide the first direct experimental evidence on the relationship between glycan structure, organization, and dynamics, offering a robust and versatile basis for a quantitative understanding of glycocalyx biology and physics at the molecular level.

Over recent decades, the glycocalyx, an extracellular organelle composed of a multitude of glycolipids, glycoproteins, proteoglycans, and glycoRNA, has gained considerable interest in cellular biology...

Super-resolution technique works with off-the-shelf optical microscopes.

For over 75 years, the United States has been viewed as a global leader in research and higher education. Today, that role is facing new challenges. Changes in funding structures, shifting priorities…

The glycocalyx is a complex layer of glycosylated biomolecules surrounding all cells in the human body. It is involved in the regulation of critical cellular processes such as immune response modulation, cell adhesion, and host-pathogen interactions. Despite these insights, the functional relationship between glycocalyx architecture and cellular state has remained elusive so far, mainly attributable to the structural diversity of glycocalyx constituents and their nanoscale organization. Here, we show that DNA-tagged lectin labeling and metabolic oligosaccharide engineering enables multiplexed super-resolution microscopy of glycocalyx constituents, yielding an atlas of glycocalyx architecture with nanometer resolution. Quantitative analysis of the obtained nanoscale map of glycocalyx constituents facilitates the extraction of characteristic spatial relationships that accurately report on cellular state. We demonstrate the capacity of our approach, which we term Glycan Atlassing, across cell and tissue types, ranging from cultured cell lines to primary immune cells, neurons, and primary patient tissue. Glycan Atlassing establishes a powerful strategy for investigating glycocalyx remodeling in development and disease, potentially enabling the development of new glycocalyx-centered targets in diagnosis and therapy. ### Competing Interest Statement The authors have declared no competing interest. Else Kröner-Fresenius-StiftungElse Kröner-Fresenius-Stiftung, https://ror.org/03zcxha54, 2020_EKEA.91 Deutsche ForschungsgemeinschaftDeutsche Forschungsgemeinschaft, https://ror.org/018mejw64, 529257351, 460333672, 270949263 Wilhelm Sander StiftungWilhelm Sander Stiftung, https://ror.org/02q83sc19, 2023.025.1 European Research CouncilEuropean Research Council, , 101118729 Alexander von Humboldt FoundationAlexander von Humboldt Foundation, https://ror.org/012kf4317,

An RNA-binding protein on leukemia cells provides an effective target in mouse models.

New findings about the sugary armor on the brain’s frontline cells could shed light on cognitive decline and diseases like Alzheimer’s – and open new avenues for treatment.

Disruption of mucin-domain glycoprotein expression and function in the endothelial glycocalyx are associated with ageing and Alzheimer’s disease, leading to dysregulated blood–brain barrier function.

Fougère, Grison et al. show that the ER–Golgi intermediate compartment (ERGIC) in plants is a tubulo-vesicular network marked by MEMBRIN proteins that is largely Golgi-independent. The plant ERGIC mat...

Membrane fluidity is a crucial feature in understanding cellular physiology. This protocol describes a robust pipeline for measuring plasma membrane fluidity using confocal imaging and new environment...