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Natural killer cell activation receptor NKP30 oligomerization depends on its n-glycosylation

DOI: 10.3390/cancers12071998 DOI Help

Authors: Ondřej Skořepa (Charles University) , Samuel Pazicky (Charles University) , Barbora Kalousková (Charles University) , Jan Bláha (Charles University) , Celeste Abreu (Charles University) , Tomáš Ječmen (Charles University) , Michal Rosůlek (Charles University; BIOCEV, Institute of Microbiology, The Czech Academy of Sciences) , Alexander Fish (Oncode Institute, Netherlands Cancer Institute) , Arthur Sedivy (Vienna Biocenter Core Facilities GmbH) , Karl Harlos (Wellcome Centre for Human Genetics, University of Oxford) , Jan Dohnalek (BIOCEV, Institute of Biotechnology, The Czech Academy of Sciences,) , Tereza Skálová (BIOCEV, Institute of Biotechnology, The Czech Academy of Sciences,) , Ondřej Vaněk (Charles University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Cancers , VOL 12

State: Published (Approved)
Published: July 2020
Diamond Proposal Number(s): 10627

Open Access Open Access

Abstract: NKp30 is one of the main human natural killer (NK) cell activating receptors used in directed immunotherapy. The oligomerization of the NKp30 ligand binding domain depends on the length of the C-terminal stalk region, but our structural knowledge of NKp30 oligomerization and its role in signal transduction remains limited. Moreover, ligand binding of NKp30 is affected by the presence and type of N-glycosylation. In this study, we assessed whether NKp30 oligomerization depends on its N-glycosylation. Our results show that NKp30 forms oligomers when expressed in HEK293S GnTI− cell lines with simple N-glycans. However, NKp30 was detected only as monomers after enzymatic deglycosylation. Furthermore, we characterized the interaction between NKp30 and its best-studied cognate ligand, B7-H6, with respect to glycosylation and oligomerization, and we solved the crystal structure of this complex with glycosylated NKp30, revealing a new glycosylation-induced mode of NKp30 dimerization. Overall, this study provides new insights into the structural basis of NKp30 oligomerization and explains how the stalk region and glycosylation of NKp30 affect its ligand affinity. This furthers our understanding of the molecular mechanisms involved in NK cell activation, which is crucial for the successful design of novel NK cell-based targeted immunotherapeutics.

Journal Keywords: NK cell; NKp30; B7-H6; glycosylation; oligomerization

Subject Areas: Biology and Bio-materials, Chemistry, Medicine

Instruments: B21-High Throughput SAXS , I02-Macromolecular Crystallography