Molecular cloning and characterization of the cathepsin B-like proteinase from the cotton boll worm, Helicoverpa armigera

Authors


  • *

    Present address: University Osnabrueck, FB5-Division of animal Physiology, D-49069 Osnabrueck, Germany.

    The nucleotide sequence reported in this paper has been submitted to GenBank with accession number AF222788.

Xiao-Fan Zhao, Department of Biology, School of Life Sciences, Shandong University, Jinan 250100, China. tel.: +86 531 8364620, fax: +86 531 8565610; e-mail: xfzhao@sdu.edu.cn

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 ≈ 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.

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