Biochemical and immunochemical characterization of 125I-labeled cuticle components of Haemonchus contortus.
Live Haemonchus contortus developmental stages were radioiodinated and then subjected to a stepwise extraction procedure consisting of a buffer extract (with or without detergent) to solubilize putative surface-associated antigenic macromolecules, followed by a detergent/beta-mercaptoethanol (BME) extract to solubilize putative cuticle collagen proteins. A buffer-extracted iodinated 100-kDa protein was present in the free-living, infective L3(2M) stage. This labeled protein was released during in vitro exsheathment of L3(2M) and was not present in the ecdysed second molt (2M) cuticle. In addition to the 100-kDa protein, exsheathment fluid contained a 70-kDa labeled protein that was not extracted from iodinated L3(2M) with either detergent or BME. The data suggest that these proteins are components of the specialized ring portion of the 2M cuticle that is enzymatically ruptured during ecdysis. The L3(2M) and the exsheathed third-stage larvae (L3) contained 3 labeled, BME-extracted, collagenase-sensitive proteins of 108, 88 and 53 kDa. In contrast, four detergent-extracted, collagenase-insensitive, iodinated proteins (143, 81, 58 and 30 kDa) were present in adult H. contortus. The 143-kDa protein was both glycosylated and immunogenic. All 4 adult cuticle proteins were released from the cuticle surface into culture fluids. Furthermore, a cysteine protease was secreted by adults which apparently hydrolyzed the released 81-, 58- and 30-kDa surface proteins.
Rhoads
M L
ML
Helminthic Diseases Laboratory, United States Department of Agriculture, Beltsville, MD 20705.
Fetterer
R H
RH
D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology; D002021:Buffers; D003094:Collagen / Q000276:immunology / Q000378:metabolism*; D003840:Deoxycholic Acid / Q000494:pharmacology; D006189:Haemonchus / Q000187:drug effects / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D007456:Iodine Isotopes; D008623:Mercaptoethanol / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D012995:Solubility
D000947:Antigens, Helminth; D002021:Buffers; D007456:Iodine Isotopes; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; D008623:Mercaptoethanol; D003094:Collagen