Two novel ficolin-like proteins act as pattern recognition receptors for invading pathogens in the freshwater crayfish Pacifastacus leniusculus
2011 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 11, no 11, 2249-2264 p.Article in journal (Refereed) Published
To isolate pathogen-associated molecular patterns (PAMPs)-binding molecules, the bacterium, Staphylococcus aureus was used as an affinity matrix to find bacteria binding proteins in the plasma of the freshwater crayfish, Pacifastacus leniusculus. Two new bacteria binding ficolin-like proteins (FLPs) were identified by 2-DE and MS analysis. The FLPs have a fibrinogen-related domain (FReD) in their C-terminal and a repeat region in their N-terminal region with putative structural similarities to the collagen-like domain of vertebrate ficolins and mannose binding lectins (MBLs). Phylogenetic analysis shows that the newly isolated crayfish FLP1 and FLP2 cluster separately from other FReD containing proteins. A tissue distribution study showed that the mRNA expression of FLP occurred mainly in the hematopoietic tissue (Hpt) and in the hepatopancreas. Recombinant FLPs exhibited agglutination activity of Gram-negative bacteria Escherichia coli and Aeromonas hydrophila in the presence of Ca2+. FLPs were able to bind to A. hydrophila, E.coli and S.aureus as judged by bacteria adsorption. Moreover, the FLPs may help crayfish to clear Gram-negative bacteria, but not Gram-positive bacteria which had been injected into the hemolymph. When Gram-negative bacteria coated with FLPs were incubated with Hpt cells, a lower death rate of the cells was found compared to control treatment. Our results suggest that FLPs function as pattern recognition receptors in the immune response of crayfish.
Place, publisher, year, edition, pages
2011. Vol. 11, no 11, 2249-2264 p.
Aeromonas hydrophila, Animal proteomics, Ficolin-like protein, FReD domain, Pattern recognition receptor, Staphylococcus aureus
Research subject Biology with specialization in Comparative Physiology
IdentifiersURN: urn:nbn:se:uu:diva-146611DOI: 10.1002/pmic.201000728ISI: 000291087700009PubMedID: 21598394OAI: oai:DiVA.org:uu-146611DiVA: diva2:398553