sequence analysis of earthworm hemolysins蚯蚓溶血素的序列分析.pdfVIP

Spectroscopy 18 (2004) 347–353 347 IOS Press Sequence analysis of earthworm hemolysins Simone König a,∗, Frank Wagner b , Ellen Kauschke c and Ines Eue d a Integrated Functional Genomics, IZKF University of Münster, Germany b Zoological Institute, University of Greifswald, Germany c Institute of Anatomy, University of Greifswald, Germany d In den Wolfseichen 5, Lohmar, Germany Abstract. Annelids are capable of defending themselves from pathogens and of recognizing degenerated self tissue. These reactions require specialized immune mechanisms that are effected by proteins and cellular reactions. Hemolytic proteins are the most striking humoral defense molecules in the earthworm Eisenia fetida. Beside their hemolytic activity these proteins possess agglutinating, antibacterial, cytotoxic, and clotting properties. Hemolytic proteins both from coelomocytes (CL39,41) and coelomic fluid (H1−3) of wildtype E. fetida were isolated and assigned to fetidin and lysenin using mass spectrometry and bioinformatic tools. Glycosylation was found for H 1. In silico analyses of the hemolysins revealed two hemolysin isoforms and consensus sites for N-glycosylation and peroxidases proximal heme-binding ligand. Keywords: Eisenia fetida, hemolysins, mass spectrometry, isoforms, glycosylation, hydrogen peroxide Abbreviations CF, coelomic fluid; CL, coelomocyte lysate; CL39,41, 39 and 41 kDa hemolysins isolated from coelo- mocyte lysate; H1−3 , three hemolytic CF proteins; LRP, lysenin-related protein; MS, mass spectrometry. 1. Introduction Earthworms, and, in fact all annelids, are interesting immunological study objects. They are capable of defending themselves from pathogens and of recognizing degenerated self tissue. These reactions re- quire specialized immunological mechanisms. Hemolytic proteins are the most striking humoral defense molecules in the earthworm