Identification and cloning of an invertebrate-type lysozyme from Eisenia andrei.
Lysozyme is a widely distributed antimicrobial protein having specificity for cleaving the beta-(1,4)-glycosidic bond between N-acetylmuramic acid (NAM) and N-acetylglucosamine (GlcNAc) of peptidoglycan of the bacterial cell walls and thus efficiently contributes to protection against infections caused mainly by Gram-positive bacteria. In the present study, we assembled a full-length cDNA of a novel invertebrate-type lysozyme from Eisenia andrei earthworm (EALys) by RT-PCR and RACE system. The primary structure of EALys shares high homology with other invertebrate lysozymes; however the highest, 72% identity, was shown for the destabilase I isolated from medicinal leech. Recombinant EALys expressed in Escherichia coli exhibited the lysozyme and isopeptidase activity. Moreover, real-time PCR revealed increased levels of lysozyme mRNA in coelomocytes of E. andrei after the challenge with both Gram-positive and Gram-negative bacteria.
Josková
Radka
R
Department of Immunology, Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
Silerová
Marcela
M
Procházková
Petra
P
Bilej
Martin
M
D000818:Animals; D001412:Bacillus subtilis / Q000276:immunology* / Q000472:pathogenicity; D001422:Bacterial Adhesion; D019759:Carbon-Nitrogen Lyases / Q000378:metabolism; D002688:Chitinases / Q000378:metabolism; D003001:Cloning, Molecular; D004447:Echinodermata / Q000235:genetics; D010450:Endopeptidases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D004927:Escherichia coli Infections / Q000276:immunology; D005944:Glucosamine / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism; D016908:Gram-Positive Bacterial Infections / Q000276:immunology; D047709:Hirudo medicinalis / Q000235:genetics; D054884:Host-Pathogen Interactions; D006868:Hydrolysis; D009112:Muramic Acids / Q000276:immunology / Q000378:metabolism; D009113:Muramidase / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D009835:Oligochaeta / Q000201:enzymology* / Q000235:genetics* / Q000276:immunology; D017385:Sequence Homology; D014774:Virulence
D009112:Muramic Acids; C031651:N-acetylmuramic acid; C098108:N-acetylglucopyranosylamine; D002688:Chitinases; D009113:Muramidase; D010450:Endopeptidases; C045348:fibrin destabilase; D019759:Carbon-Nitrogen Lyases; C034300:isopeptidase; D005944:Glucosamine