Natural Killer Cell Activation Receptor NKp30 Oligomerization Depends on Its N-Glycosylation
Open Access
- 20 July 2020
- Vol. 12 (7), 1998
- https://doi.org/10.3390/cancers12071998
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.Funding Information
- European Regional Development Fund (CZ.1.05/1.1.00/02.0109)
- Univerzita Karlova v Praze (SVV 260427/2020)
- Wellcome Trust (203141/Z/16/Z)
- European Cooperation in Science and Technology (CA15126)
This publication has 61 references indexed in Scilit:
- Crystal structure of human natural cytotoxicity receptor NKp30 and identification of its ligand binding siteProceedings of the National Academy of Sciences of the United States of America, 2011
- Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6The Journal of Experimental Medicine, 2011
- XDSActa crystallographica. Section D, Structural biology, 2010
- MolProbity: all-atom structure validation for macromolecular crystallographyActa crystallographica. Section D, Structural biology, 2009
- The B7 family member B7-H6 is a tumor cell ligand for the activating natural killer cell receptor NKp30 in humansThe Journal of Experimental Medicine, 2009
- Inference of Macromolecular Assemblies from Crystalline StateJournal of Molecular Biology, 2007
- Phasercrystallographic softwareJournal of Applied Crystallography, 2007
- Inhibition of the NKp30 activating receptor by pp65 of human cytomegalovirusNature Immunology, 2005
- UCSF Chimera?A visualization system for exploratory research and analysisJournal of Computational Chemistry, 2004
- Size-Distribution Analysis of Macromolecules by Sedimentation Velocity Ultracentrifugation and Lamm Equation ModelingBiophysical Journal, 2000