Mosquito cathepsin B-like protease involved in embryonic degradation of vitellin is produced as a latent extraovarian precursor.
Here we report identification of a novel member of the thiol protease superfamily in the yellow fever mosquito, Aedes aegypti. It is synthesized and secreted as a latent proenzyme in a sex-, stage-, and tissue-specific manner by the fat body, an insect metabolic tissue, of female mosquitoes during vitellogenesis in response to blood feeding. The secreted, hemolymph form of the enzyme is a large molecule, likely a hexamer, consisting of 44-kDa subunits. The deduced amino acid sequence of this 44-kDa precursor shares high similarity with cathepsin B but not with other mammalian cathepsins. We have named this mosquito enzyme vitellogenic cathepsin B (VCB). VCB decreases to 42 kDa after internalization by oocytes. In mature yolk bodies, VCB is located in the matrix surrounding the crystalline yolk protein, vitellin. At the onset of embryogenesis, VCB is further processed to 33 kDa. The embryo extract containing the 33-kDa VCB is active toward benzoyloxycarbonyl-Arg-Arg-para-nitroanilide, a cathepsin B-specific substrate, and degrades vitellogenin, the vitellin precursor. Both of these enzymatic activities are prevented by trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64), a thiol protease inhibitor. Furthermore, addition of the anti-VCB antibody to the embryonic extract prevented cleavage of vitellogenin, strongly indicating that the activated VCB is involved in embryonic degradation of vitellin.
Cho
W L
WL
Department of Parasitology, National Yang-Ming University, Taipei 112, Taiwan.
Tsao
S M
SM
Hays
A R
AR
Walter
R
R
Chen
J S
JS
Snigirevskaya
E S
ES
Raikhel
A S
AS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D009033:Culicidae / Q000196:embryology / Q000201:enzymology*; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary; D004527:Egg Proteins / Q000378:metabolism*; D004705:Endocytosis; D004792:Enzyme Precursors / Q000096:biosynthesis*; D005260:Female; D006868:Hydrolysis; D007150:Immunohistochemistry; D007814:Larva / Q000378:metabolism*; D008969:Molecular Sequence Data; D010053:Ovary / Q000201:enzymology / Q000648:ultrastructure; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D004527:Egg Proteins; D004792:Enzyme Precursors; D003546:Cysteine Endopeptidases