N-Glycosylation-dependent regulation of immune-specific tetraspanins CD37 and CD53

Sjoerd van Deventer; Ilse A Hoogvliet; Merel van de Voort; Frank Arnold; Martin Ter Beest; Annemiek B van Spriel

doi:10.1016/j.bpj.2023.11.3399

N-Glycosylation-dependent regulation of immune-specific tetraspanins CD37 and CD53

Biophys J. 2023 Nov 28:S0006-3495(23)04100-0. doi: 10.1016/j.bpj.2023.11.3399. Online ahead of print.

Authors

Sjoerd van Deventer¹, Ilse A Hoogvliet¹, Merel van de Voort¹, Frank Arnold², Martin Ter Beest¹, Annemiek B van Spriel³

Affiliations

¹ Department of Medical BioSciences, Radboud University Medical Center, Nijmegen, the Netherlands.
² Cancer Biology and Genetics Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, New York.
³ Department of Medical BioSciences, Radboud University Medical Center, Nijmegen, the Netherlands. Electronic address: annemiek.vanspriel@radboudumc.nl.

PMID: 38031400
DOI: 10.1016/j.bpj.2023.11.3399

Abstract

Tetraspanin proteins play an important role in many cellular processes as they are key organizers of different receptors on the plasma membrane. Most tetraspanins are highly glycosylated at their large extracellular loop; however, little is known about the function of tetraspanin glycosylation in immune cells. In this study we investigated the effects of glycosylation of CD37 and CD53, two tetraspanins important for cellular and humoral immunity. Broad and cell-specific repertoires of N-glycosylated CD37 and CD53 were observed in human B cells. We generated different glycosylation mutants of CD37 and CD53 and analyzed their localization, nanoscale plasma membrane organization, and partner protein interaction capacity. Abrogation of glycosylation in CD37 revealed the importance of this modification for CD37 surface expression, whereas surface expression of CD53 was unaffected by its glycosylation. Single-molecule dSTORM microscopy revealed that the nanoscale organization of CD53 was not dependent on glycosylation. CD37 interaction with its partner proteins CD53 and CD20 was affected by glycosylation in a localization-dependent way, whereas its interaction with IL-6Rα was independent of glycosylation. Surprisingly, glycosylation was found to inhibit the interaction between CD53 and its partner proteins CD45, CD20, and, to a lesser extent CD37. Together, our data show that glycosylation affects the interaction capacity of immune-specific tetraspanins CD37 and CD53, which adds another layer of regulation to immune membrane organization.