Proteomic analysis of the Drosophila larval hemolymph clot.
Components of the insect clot, an extremely rapid forming and critical part of insect immunity, are just beginning to be identified (1). Here we present a proteomic comparison of larval hemolymph before and after clotting to learn more about this process. This approach was supplemented by the identification of substrates for the enzyme transglutaminase, which plays a role in both vertebrate blood clotting (as factor XIIIa) and hemolymph coagulation in arthropods. Hemolymph proteins present in lower amounts after clotting include CG8502 (a protein with a mucin-type domain and a domain with similarity to cuticular components), CG11313 (a protein with similarity to prophenoloxidase-activating proteases), and two phenoloxidases, lipophorin, a secreted gelsolin, and CG15825, which had previously been isolated from clots (2). Proteins whose levels increase after clotting include a ferritin-subunit and two members of the immunoglobulin family with a high similarity to the small immunoglobulin-like molecules involved in mammalian innate immunity. Our results correlate with findings from another study of coagulation (2) that involved a different experimental approach. Proteomics allows the isolation of novel candidate clotting factors, leading to a more complete picture of clotting. In addition, our two-dimensional protein map of cell-free Drosophila hemolymph includes many additional proteins that were not found in studies performed on whole hemolymph.
Karlsson
Christine
C
Department of Molecular Biology and Functional Genomics, University of Stockholm, 10691 Stockholm, Sweden.
Korayem
Ahmed M
AM
Scherfer
Christoph
C
Loseva
Olga
O
Dushay
Mitchell S
MS
Theopold
Ulrich
U
D000595:Amino Acid Sequence; D000818:Animals; D001798:Blood Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology; D004330:Drosophila / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D029721:Drosophila Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology; D015180:Electrophoresis, Gel, Two-Dimensional; D017344:Genes, Insect; D006458:Hemolymph / Q000276:immunology / Q000502:physiology*; D007814:Larva / Q000502:physiology; D008969:Molecular Sequence Data; D040901:Proteomics; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism
D001798:Blood Proteins; D029721:Drosophila Proteins; C512855:fon protein, Drosophila; D011503:Transglutaminases