Rat natural killer cell antigen, NKR-P1, related to C-type animal lectins is a carbohydrate-binding protein

Karel Bezouška, Georgios Vlahas, Ondrej Horváth, Gabriela Jinochová, Anna Fišerová, Roberto Giorda, William H. Chambers, Ten Feizi, Miloslav Pospíšil

Research output: Contribution to journalArticle

Abstract

Natural killer receptor protein 1 (NKR-P1, a family of proteins), which is a dimeric transmembrane protein predominantly on rat and murine natural killer cells, contains an extracellular motif related to calcium-dependent animal lectins. The domain architecture of this protein and the finding that its cross-linking with antibody results in activation of natural killer cells make it a promising candidate for a receptor function. We have expressed a full-length NKR-P1 protein of the rat in COS cells and prepared soluble extracellular fragments by controlled proteolysis or by expression of truncated cDNA in bacteria. Dimerization of soluble NKR-P1 is predominantly dependent on the presence of an intact juxtamembrane stalk region and independent of N-glycosylation. Binding and inhibition studies using monosaccharides and neoglycoconjugates indicate that NKR-P1 is a lectin with a preference order of GalNAc > GlcNAc >> Fuc >> Gal > Man. At neutral pH, Ca2+ is tightly associated with the protein such that only a proportion can be removed by 10 mM EGTA. However, NKR-P1 can be decalcified completely at pH 10 with a total loss of carbohydrate binding. After recalcification at pH 8, carbohydrate binding is completely restored. Thus, NKR-P1 differs from other calcium-dependent animal lectins investigated so far in its pattern of monosaccharide recognition and in the tightness of Ca2+ binding.

Original languageEnglish
Pages (from-to)16945-16952
Number of pages8
JournalJournal of Biological Chemistry
Volume269
Issue number24
Publication statusPublished - Jun 17 1994

ASJC Scopus subject areas

  • Biochemistry

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    Bezouška, K., Vlahas, G., Horváth, O., Jinochová, G., Fišerová, A., Giorda, R., Chambers, W. H., Feizi, T., & Pospíšil, M. (1994). Rat natural killer cell antigen, NKR-P1, related to C-type animal lectins is a carbohydrate-binding protein. Journal of Biological Chemistry, 269(24), 16945-16952.