Purification, cloning, and expression of a novel salivary anticomplement protein from the tick, Ixodes scapularis.
The alternative pathway of complement is an important defense against pathogens and in tick rejection reactions. The tick Ixodes scapularis is able to feed repeatedly on its natural host and has a salivary anticomplement activity that presumably facilitates feeding. In this study, we purified and then obtained the amino-terminal sequence of the I. scapularis salivary anticomplement (Isac). We found a full-length clone coding for Isac by random screening of a salivary gland cDNA library. Expressing Isac cDNA in COS cells reproduced the activity found in tick saliva, namely, inhibition of rabbit erythrocyte lysis by human serum in the presence of Mg(2+) and EGTA, inhibition of C3b binding to agarose in the presence of Mg(2+) and EGTA, and acceleration of factor Bb uncoupling from the C3 convertase generated by the alternative pathway. Recombinant Isac had no effect on the recalcification time of human platelet-poor plasma or in the classical complement pathway, indicating that it is a specific inhibitor similar to the regulators of complement activation of the alternative pathway such as factor H. Isac, however, has no similarity to any protein in the GenBank(TM) data base, indicating that it is a novel and relatively small (18.5 kDa) anticomplement molecule.
Valenzuela
J G
JG
NIAID, National Institutes of Health, Laboratory of Parasitic Diseases, Section of Medical Entomology, Bethesda, Maryland 20892-0425, USA.
Charlab
R
R
Mather
T N
TN
Ribeiro
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D011817:Rabbits; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D013987:Ticks / Q000737:chemistry*
D018076:DNA, Complementary; C411881:Isac protein, Ixodes scapularis; D012471:Salivary Proteins and Peptides