Molecular cloning of cDNA for Sarcophaga prolyl endopeptidase and characterization of the recombinant enzyme produced by an E. coli expression system

Sumio Ohtsuki, Ko Ichi Homma, Shoichiro Kurata, Shunji Natori

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

A cDNA for prolyl endopeptidase (PEP) of Sarcophaga peregrina (flesh fly) was cloned and its sequence determined. The overall amino acid sequence identity between Sarcophaga and mammalian PEPs was 53%, indicating that these enzymes are structurally very similar. Northern blot hybridization revealed that the Sarcophaga PEP gene was activated significantly at the eversion stage of imaginal disc differentiation. We obtained recombinant PEP by expressing the cDNA in Escherichia coli. The recombinant and authentic enzymes showed almost identical characteristics, in terms of substrate specificities and sensitivities to inhibitors.

Original languageEnglish
Pages (from-to)337-343
Number of pages7
JournalInsect Biochemistry and Molecular Biology
Volume27
Issue number4
DOIs
Publication statusPublished - 1997 Jan 1
Externally publishedYes

Keywords

  • Imaginal disc differentiation
  • Prolyl endopeptidase
  • Recombinant enzyme
  • Sarcophaga peregrina

ASJC Scopus subject areas

  • Insect Science
  • Biochemistry

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