title: Purification and characterization of the western spruce budworm larval midgut proteinases and comparison of gut activities of laboratory-reared and field-collected insects.
abstract: Three proteolytic enzymes, trypsin, chymotrypsin, and aminopeptidase-N (APN), were purified from laboratory-reared western spruce budworm, Choristoneura occidentalis [Freeman], larvae. Budworm trypsin exhibited a high degree of substrate specificity, was inactivated by DFP and TLCK, and was inhibited by trypsin inhibitors. The western spruce budworm chymotrypsin hydrolyzed SAAPFpNA and SAAPLpNA, but not SFpNA, SGGFpNA, SGGLpNA or BTpNA. The chymotrypsin was inactivated by DFP, and was inhibited by chymostatin and the chymotrypsin inhibitor, POT-1. Purified budworm chymotrypsin exhibited little BTEE esterolytic activity and was insensitive to inhibition with TPCK. The N-terminal sequence of budworm trypsin, chymotrypsin, and APN were obtained. Similar levels of trypsin and APN gut activities were found in laboratory-reared and field-collected larvae. However, in comparison to laboratory-reared insects, considerably less chymotrypsin activity, and a much higher level of gut carboxypeptidase activity were found in field-collected western spruce budworm larvae.
authors.affiliation: USDA Forest Service, Northeastern Experiment Station, Delaware, OH 43015-8640, USA. avalaiti/ne_de@fs.fed.us
authors.forename: A P
authors.initials: AP
authors.lastname: Valaitis
authors.forename: S
authors.initials: S
authors.lastname: Augustin
authors.forename: K M
authors.initials: KM
authors.lastname: Clancy
mesh_terms: D000818:Animals; D018826:CD13 Antigens / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D004064:Digestive System / Q000201:enzymology; D004791:Enzyme Inhibitors; D007814:Larva; D009036:Moths / Q000201:enzymology*; D014357:Trypsin / Q000302:isolation & purification / Q000378:metabolism*; D014361:Trypsin Inhibitors
chemical_list: D004791:Enzyme Inhibitors; D014361:Trypsin Inhibitors; D018826:CD13 Antigens; D002918:Chymotrypsin; D014357:Trypsin