Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 108, Issue 34, Pages 14270-14275Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1111415108
Keywords
pattern-recognition receptors; sortase A; polysaccharide
Categories
Funding
- National Institutes of Health [GM040266]
- National Institute of Allergy and Infectious Diseases [5 R01 AI087879]
- Fulbright/Spanish Ministry of Education and Science [FU2006-0983]
- National Institutes of Health National Research Service [F32 AI729353]
- Margaret and Herman Sokol Fellowship in Biomedical Research
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Dectin-1, the major beta-glucan receptor in leukocytes, triggers an effective immune response upon fungal recognition. Here we use sortase-mediated transpeptidation, a technique that allows placement of a variety of probes on a polypeptide backbone, to monitor the behavior of labeled functional dectin-1 in live cells with and without fungal challenge. Installation of probes on dectin-1 by sortagging permitted highly specific visualization of functional protein on the cell surface and its subsequent internalization upon ligand presentation. Retrieval of sortagged dectin-1 expressed in macrophages uncovered a unique interaction between dectin-1 and galectin-3 that functions in the proinflammatory response of macrophages to pathogenic fungi. When macrophages expressing dectin-1 are exposed to Candida albicans mutants with increased exposure of beta-glucan, the loss of galectin-3 dramatically accentuates the failure to trigger an appropriate TNF-alpha response.
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