title: Molecular cloning and characterization of the cathepsin B-like proteinase from the cotton boll worm, Helicoverpa armigera.
abstract: An enzyme purified from the ovaries of Helicoverpa armigera, as an active form with molecular mass of 30 kDa on SDS-PAGE, was identified as a cysteine proteinase because it could be inhibited by E-64, a specific inhibitor of cysteine proteinase, and required reducing conditions for activity. This enzyme was further identified as a cathepsin B-like cysteine proteinase by partial amino acid sequencing. A cDNA encoding this proteinase was cloned from H. armigera, using degenerate primers and RACE techniques. Results of Northern blots indicated that the mRNA encoding the proteinase was transcribed in the ovaries, the fat bodies of female and male adults, pupae and in the larvae. No mRNA was detected from the larval epidermis or from the midgut. Hence, transcription of the cathepsin B-like cysteine proteinase from H. armigera was tissue-specific, but not gender- or developmental stage-specific. However, proteolytic activities were only detected from ovaries, and adult female and male fat bodies. No activity was observed from pupal and larval fat bodies, from the larval epidermis or from the midgut. Only one form of mRNA of approximately 1100 bases was detected, and in situ hybridization showed that the transcripts were distributed in the adult female fat bodies, follicular cells and the oocytes. Since the proteinase expressed in ovaries was able to degrade vitellin in vitro, it may be involved in the degradation of vitellin during embryonic development.
authors.affiliation: Department of Biology, School of Life Sciences, Shandong University, Jinan 250100, China.
authors.forename: X-F
authors.initials: XF
authors.lastname: Zhao
authors.forename: J-X
authors.initials: JX
authors.lastname: Wang
authors.forename: X-L
authors.initials: XL
authors.lastname: Xu
authors.forename: R
authors.initials: R
authors.lastname: Schmid
authors.forename: H
authors.initials: H
authors.lastname: Wieczorek
mesh_terms: D000273:Adipose Tissue / Q000201:enzymology; D000595:Amino Acid Sequence; D000818:Animals; D002401:Cathepsin B / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005260:Female; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D010802:Phylogeny; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers; D010450:Endopeptidases; D002401:Cathepsin B