Structural basis for the recognition of C20: 2-αGalCer by the invariant natural killer T cell receptor-like antibody L363

Esther Dawen Yu, Enrico Girardi, Jing Wang, Thien Thi Mac, Karl O A Yu, Serge Van Calenbergh, Steven A. Porcelli, Dirk M. Zajonc

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Natural killer T (NKT) cells express a semi-invariant Vα14 T cell receptor (TCR) and recognize structurally diverse antigens presented by the antigen-presenting molecule CD1d that range from phosphoglycerolipids to α- and β-anomeric glycosphingolipids, as well as microbial α-glycosyl diacylglycerolipids. Recently developed antibodies that are specific for the complex of the prototypical invariant NKT (iNKT) cell antigen αGalCer (KRN7000) bound to mouse CD1d have become valuable tools in elucidating the mechanism of antigen loading and presentation. Here, we report the 3.1 Å resolution crystal structure of the Fab of one of these antibodies, L363, bound to mCD1d complexed with the αGalCer analog C20:2, revealing that L363 is an iNKT TCR-like antibody that binds CD1d-presented αGalCer in a manner similar to the TCR. The structure reveals that L363 depends on both the L and H chains for binding to the glycolipid-mCD1d complex, although only the L chain is involved in contacts with the glycolipid antigen. The H chain of L363 features residue Trp-104, which mimics the TCR CDR3α residue Leu-99, which is crucial for CD1d binding. We characterized the antigen-specificity of L363 toward several different glycolipids, demonstrating that whereas the TCR can induce structural changes in both antigen and CD1d to recognize disparate lipid antigens, the antibody L363 can only induce the F′ roof formation in CD1d but fails to reorient the glycolipid headgroup necessary for binding. In summary, L363 is a powerful tool to study mechanism of iNKT cell activation for structural analogs of KRN7000, and our study can aid in the design of antibodies with altered antigen specificity.

Original languageEnglish (US)
Pages (from-to)1269-1278
Number of pages10
JournalJournal of Biological Chemistry
Volume287
Issue number2
DOIs
StatePublished - Jan 6 2012

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Natural Killer Cell Receptors
Natural Killer T-Cells
T-Cell Antigen Receptor
Glycolipids
Antigens
CD1d Antigen
Antibodies
T-cells
Glycosphingolipids
Viral Tumor Antigens
Antigen Presentation
Roofs
Lipids
Crystal structure
Chemical activation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Yu, E. D., Girardi, E., Wang, J., Mac, T. T., Yu, K. O. A., Van Calenbergh, S., ... Zajonc, D. M. (2012). Structural basis for the recognition of C20: 2-αGalCer by the invariant natural killer T cell receptor-like antibody L363. Journal of Biological Chemistry, 287(2), 1269-1278. https://doi.org/10.1074/jbc.M111.308783

Structural basis for the recognition of C20 : 2-αGalCer by the invariant natural killer T cell receptor-like antibody L363. / Yu, Esther Dawen; Girardi, Enrico; Wang, Jing; Mac, Thien Thi; Yu, Karl O A; Van Calenbergh, Serge; Porcelli, Steven A.; Zajonc, Dirk M.

In: Journal of Biological Chemistry, Vol. 287, No. 2, 06.01.2012, p. 1269-1278.

Research output: Contribution to journalArticle

Yu, Esther Dawen ; Girardi, Enrico ; Wang, Jing ; Mac, Thien Thi ; Yu, Karl O A ; Van Calenbergh, Serge ; Porcelli, Steven A. ; Zajonc, Dirk M. / Structural basis for the recognition of C20 : 2-αGalCer by the invariant natural killer T cell receptor-like antibody L363. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 2. pp. 1269-1278.
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