Glycan-Based Connectivity Regulates the Hierarchical Organization of Membrane Receptors by Coupling their Micro- and Nano-Scale Lateral Mobility

Abstract

Glycan-protein interactions are emerging as important modulators of membrane protein organization and dynamics, regulating multiple cellular functions. In particular, it has been postulated that glycan-mediated interactions regulate surface residence time of glycoproteins and endocytosis. How this precisely occurs is poorly understood. We applied a combination of super-resolution nanoscopy and single molecule-based approaches to study the role of glycan-based interactions on the dynamics of the glycosylated pathogen recognition receptor DC-SIGN, at the nano- and micrometer scale. We find that cell surface glycan-mediated interactions do not influence the nanoscale lateral organization of DC-SIGN in nanoclusters but restrict the mobility of the receptor to distinct micron-size membrane regions. These meso-scale regions are in turn enriched by the endocytic protein clathrin, thereby dynamically promoting DC-SIGN transient nano-scale arrest and interaction with clathrin. Disruption of glycan-based connectivity leads to larger membrane exploration, reduced clathrin interaction and compromised clathrin-dependent internalization of virus-like particles. Therefore, our work uncovers a novel mechanism through which glycan-protein interactions act as decision-makers in fine-tuning membrane-related functions by dynamically coupling micro- and nanoscale receptor lateral mobility, thus adding a new layer of regulation to the hierarchical spatiotemporal organization of the cell membrane.