Unlike Natural Killer (NK) p30, Natural Cytotoxicity Receptor NKp44 Binds to Multimeric α2,3-NeuNAc-Containing N-Glycans
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概要
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Natural cytotoxicity receptor 2 (NCR2 or natural killer (NK)p44) and NCR3 (NKp30) bind to heparin and heparin sulfate; however, other natural ligands have yet to be identified. We previously reported that NCR1 (NKp46) can bind to multimeric NeuNAc-containing N-glycans and sulfated glycans. In this study, we investigated whether NKp44 and NKp30 can bind to NeuNAc-containing glycans using their common recombinant extracellular domain tagged with 6×His (NKp44-H6 and NKp30-H6). NKp44-H6, but not NKp30-H6, bound multimeric sialyl Lewis X expressing transferrin secreted by HepG2 cells (HepTF) with a Kd of 420 nM. Competitive and direct binding assays revealed that NKp44-H6 mainly recognizes α2,3-NeuNAc residues on non-reducing ends of N-glycans on HepTF. Moreover, site-directed mutants of NKp44-H6, such as R47Q, R55Q, R92Q, R95Q, K103Q, and R106Q, had reduced binding to α2,3-sialylated N-glycans. These results suggest that NKp44 binds to α2,3-sialylated N-glycans through ionic interactions, and that these binding sites might have some overlap with heparin binding sites.
著者
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Ito Kenichiro
Department Of Applied Physics Faculty Of Science And Engineering Waseda University
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Yanai Kazuyuki
Deparetment Of Gene Diagnositics And Theraputics Resarch Institute International Medical Center Of J
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Higai Koji
Department Of Clinical Chemistry School Of Pharmaceutical Sciences Toho University
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Azuma Yutaro
Department Of Biochemistry Tokyo Dental College
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Matsumoto Kojiro
Department Of Clinical Chemistry Faculty Of Pharmaceutical Sciences Toho University
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Yanai Kazuyuki
Department of Biomolecular Science, Faculty of Science, Toho University
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Shinoda Chihiro
Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Toho University
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Sakurai Mizuha
Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Toho University
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Ito Kenichiro
Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Toho University
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