Cathepsin L function in insect moulting: molecular cloning and functional analysis in cotton bollworm, Helicoverpa armigera.
Moulting is an essential process of insect development but little is known about cysteine proteases in the process. Here, we detail a proteolytic activity profile from fifth larval instar to new pupae of the lepidopteran Helicoverpa armigera. At fifth to sixth instar moulting, the activities were significantly higher than those in non-moulting stages, and were inhibited by the cysteine protease inhibitor, 2S, 3S-trans-epoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester (E-64), or by the cathepsin L-selective inhibitor CLIK148. Further, a 1513 bp cathepsin L cDNA (Har-CL) was isolated from the H. armigera larval cuticle and epidermis layer. Har-CL gene expression, which is correlated closely with ecdysone, was higher during larval moulting. Injection of E-64 or CLIK148 resulted in delayed fifth to sixth instar moulting, suggesting an essential role for cathepsin L in larval moulting.
Liu
J
J
Department of Molecular and Cell Biology, University of Science and Technology of China, Hefei, China.
Shi
G-P
GP
Zhang
W-Q
WQ
Zhang
G-R
GR
Xu
W-H
WH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D056668:Cathepsin L; D002403:Cathepsins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000302:isolation & purification; D004440:Ecdysone / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004817:Epidermis / Q000187:drug effects; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D003368:Gossypium / Q000469:parasitology*; D006458:Hemolymph / Q000187:drug effects / Q000378:metabolism; D007814:Larva / Q000187:drug effects; D007915:Lepidoptera / Q000254:growth & development / Q000502:physiology*; D008018:Life Cycle Stages; D008969:Molecular Sequence Data; D018978:Molting / Q000187:drug effects / Q000502:physiology*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D004440:Ecdysone; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056668:Cathepsin L