Binding of porcine ficolin-alpha to lipopolysaccharides from Gram-negative bacteria and lipoteichoic acids from Gram-positive bacteria.
Protein(s) reactive with N-acetyl-D-glucosamine (GlcNAc) was isolated from porcine nonimmune serum. The molecular weight of the purified protein was found to be mainly 40 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. The N-terminal 10 amino acid sequence of the purified protein were found to be identical to that of porcine ficolin-alpha reported previously. In enzyme-linked immunosorbent assay, the purified protein was found to react with lipopolysaccharides (LPS) from different Gram-negative bacteria such as Esherichia coli, Salmonella typhimurium, Salmonella enteritidis, Salmonella abortus equi, Pseudomonas aeruginosa, Shigella flexeneri, and Serratia marcescens and with lipoteichoic acid (LTA) from Gram-positive bacteria such as Streptococcus sanguis, Bacillus subtilis, Streptococcus pyogenes, and Staphylococcus aureus. The purified protein also reacted with E. coli O26 isolated from food poisoning and bovine feces and heat-treated Gram-positive bacteria such as S. aureus, B. cereus, B. subtilis, Enterococcus faecium, and Corynebacterum bovis. On the other hand, porcine IgG isolated from nonimmune serum showed different reactivity with these LPS, LTA, and heat-treated bacterial cells. From the present findings, purified porcine serum protein reactive with GlcNAc is concluded to be ficolin-alpha playing an important role(s) in innate immunity against microbial infection with Gram-positive and -negative bacteria.
Nahid
Abu M
AM
Laboratory of Veterinary Microbiology, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan.
Sugii
Shunji
S
D000595:Amino Acid Sequence; D000818:Animals; D006090:Gram-Negative Bacteria / Q000378:metabolism*; D006094:Gram-Positive Bacteria / Q000378:metabolism*; D006358:Hot Temperature; D007074:Immunoglobulin G / Q000276:immunology; D037102:Lectins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D013552:Swine / Q000378:metabolism*; D000097604:Ficolins
D007074:Immunoglobulin G; D037102:Lectins; D008070:Lipopolysaccharides; D000097604:Ficolins