Distinct carbohydrate recognition domains of an invertebrate defense molecule recognize Gram-negative and Gram-positive bacteria
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
11585829
DOI
10.1074/jbc.m107220200
PII: S0021-9258(19)37373-9
Knihovny.cz E-resources
- MeSH
- Enzyme Activation MeSH
- Cytotoxins chemistry metabolism MeSH
- DNA Primers MeSH
- Gram-Negative Bacteria metabolism MeSH
- Gram-Positive Bacteria metabolism MeSH
- Lectins * MeSH
- Carbohydrate Metabolism * MeSH
- Molecular Sequence Data MeSH
- Oligochaeta enzymology metabolism MeSH
- Amino Acid Sequence MeSH
- Base Sequence MeSH
- Monophenol Monooxygenase metabolism MeSH
- Binding Sites MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- coelomic cytolytic factor 1, Eisenia fetida MeSH Browser
- Cytotoxins MeSH
- DNA Primers MeSH
- Lectins * MeSH
- Monophenol Monooxygenase MeSH
Coelomic fluid of Eisenia foetida earthworms (Oligochaeta, Annelida) contains a 42-kDa defense molecule named CCF for coelomic cytolytic factor. By binding microbial antigens, namely the O-antigen of lipopolysaccharide (LPS), beta-1,3-glucans, or N,N'-diacetylchitobiose present, respectively, on Gram-negative bacteria or yeast cell walls, CCF triggers the prophenoloxidase activating pathway. We report that CCF recognizes lysozyme-predigested Gram-positive bacteria or the peptidoglycan constituent muramyl dipeptide as well as muramic acid. To identify the pattern recognition domains of CCF, deletion mutants were tested for their ability to reconstitute the prophenoloxidase cascade in E. foetida coelomic fluid depleted of endogenous CCF in the presence of LPS, beta-1,3-glucans, N,N'-diacetylchitobiose, and muramic acid. In addition, affinity chromatography of CCF peptides was performed on immobilized beta-1,3-glucans or N,N'-diacetylchitobiose. We found that the broad specificity of CCF for pathogen-associated molecular patterns results from the presence of two distinct pattern recognition domains. One domain, which shows homology with the polysaccharide and glucanase motifs of beta-1,3-glucanases and invertebrate defense molecules located in the central part of the CCF polypeptide chain, interacts with LPS and beta-1,3-glucans. The C-terminal tryptophan-rich domain mediates interactions of CCF with N,N'-diacetylchitobiose and muramic acid. These data provide evidence for the presence of spatially distinct carbohydrate recognition domains within this invertebrate defense molecule.
References provided by Crossref.org
Evidence for proteins involved in prophenoloxidase cascade Eisenia fetida earthworms
GENBANK
AF030028, AF395805
