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Papers In Press, published online ahead of print May 15, 2004
Biochemistry Dept., University of Oxford, Oxford OX1 3QU
Corresponding Author: Palaniyar.Nadesalingam{at}bioch.ox.ac.uk
Collectins are a family of innate immune proteins that contain fibrillar collagen-like regions and globular carbohydrate recognition domains (CRDs). The CRDs of these proteins recognize various microbial surface-specific carbohydrate patterns, particularly hexoses. We hypothesized that collectins such as pulmonary surfactant proteins (SP-) A and D, and serum protein mannose-binding lectin (MBL), could recognize nucleic acids, pentose-based anionic phosphate polymers. Here, we show that collectins bind DNA from a variety of origins, including bacteria, mice and synthetic oligonucleotides. Pentoses such as arabinose, ribose and deoxyribose inhibit the interaction between SP-D and mannan, one of the well-studied hexose ligands for SP-D, and biologically relevant D-forms of the pentoses are better competitors than the L-forms. In addition, DNA and RNA polymer-related compounds, such as nucleotide di- and triphosphates, also inhibit the carbohydrate binding ability of SP-D, or ~60 kDa trimeric recombinant fragments of SP-D that are composed of the alpha-helical coiled-coil neck region and 3 CRDs (SP-D(n/CRD)), or SP-D(n/CRD) with 8 Gly-X-Y repeats (SP-D(GXY)8(n/CRD)). Direct binding and competition studies suggest that collectins bind nucleic acid via their CRDs as well as via their collagen-like regions, and that SP-D binds DNA more effectively than do SP-A and MBL at physiological salt conditions. Furthermore, the SP-D(GXY)8(n/CRD) fragment co-localize with DNA, and the protein competes the interaction between propidium iodide, a DNA-binding dye, and apoptotic cells. In conclusion, we show that collectins are a new class of proteins that bind free DNA and DNA present on apoptotic cells via both of their globular CRDs and collagen-like regions. Collectins may therefore play an important role in decreasing the inflammation caused by DNA in lungs and other tissues.
J. Biol. Chem, 10.1074/jbc.M403763200
Submitted on April 5, 2004
Revised on April 27, 2004
Accepted on May 14, 2004
Nucleic acid is a novel ligand for innate immune pattern recognition collectins surfactant proteins A and D and mannose-binding lectin
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