Characterization of hemolytic and cytotoxic Gallysins: a relationship with arylphorins.
Gallysin-1, an inducible effector protein in the protective response of Galleria mellonella larvae is a 75 kDa component of hemolytically active material (HAM) isolated from immune cell-free hemolymph. The sequence of the first 20 N-terminal amino acids of the antibacterial protein Gallysin-1 is identical to the predicted sequence of the first 20 amino acids of the Galleria arylphorin Lhp76 (larval hemolymph protein 76). A murine monoclonal antibody to the 20 amino acid N-terminal peptide of Gallysin-1 (GYPQYHYDVETRKLDPSLVN) provides additional evidence for a link between Gallysin-1 and Lhp76, and is used to characterize HAM further. HAM, initially characterized as a mixture of two proteins, Gallysin-1 and a 69 kDa component is now identified as a 450-500 kDa heteromultimer, designated Gallysin. In vivo levels of Gallysin rise during the effector phase of an induced immune response. The monoclonal antibody inhibits the hemolytic activity of Gallysin. In addition to a hemolytic activity for mammalian erythrocytes, Gallysin possesses a cytotoxic activity for the human tumor cell line, K562. Lipopolysaccharides (LPS) and a Pseudomonas aeruginosa vaccine induce a cytotoxic activity which reaches its maximum levels in the hemolymph early (2 hours post-vaccination) in the protective response. The partially purified cytotoxic material (Cyt-M) obtained from cell-free hemolymph collected 2 hours after vaccination has hemolytic activity and shows structural similarities to Gallysin and Lhp76. The previously established role of Gallysin-1 as an effector protein in the protective response of Galleria mellonella indicates that arylphorins may play a role in insect immune responses.
Beresford
P J
PJ
Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada.
Basinski-Gray
J M
JM
Chiu
J K
JK
Chadwick
J S
JS
Aston
W P
WP
D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology; D001704:Biopolymers / Q000737:chemistry / Q000276:immunology; D002474:Cell-Free System / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000502:physiology; D006458:Hemolymph / Q000737:chemistry / Q000187:drug effects / Q000276:immunology; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000276:immunology / Q000502:physiology; D006461:Hemolysis / Q000187:drug effects / Q000276:immunology; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology; D007938:Leukemia; D008969:Molecular Sequence Data; D009036:Moths; D011817:Rabbits; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D000890:Anti-Infective Agents; D000911:Antibodies, Monoclonal; D001704:Biopolymers; D006023:Glycoproteins; D006460:Hemolysin Proteins; D019476:Insect Proteins; C041137:storage proteins, Insecta