Characterization of a blood activated chitinolytic system in the midgut of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi

J. M. Ramalho-Ortigão, S. Kamhawi, M. B. Joshi, David Reynoso, P. G. Lawyer, D. M. Dwyer, D. L. Sacks, J. G. Valenzuela

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

We characterized a cDNA from Phlebotomus papatasi, PpChit1, which encodes a midgut specific chitinase and show the presence of a functional, blood-induced chitinolytic system in sand flies. PpChit1 is detected only in the midgut and is regulated by blood feeding. A recombinant protein (rPpChit1) produced in HEK 293-F cells exhibited a similar activity profile to that found in the native protein against several specific substrates, including an oligomeric glycol chitin and synthetic 4-methyl-umbelliferone labelled substrates. Western blotting showed that the native protein is recognized by mouse polyclonal antibodies against rPpChit1. Additionally, the rPpChit1 and the native chitinase displayed similar retention times in a HPLC size fractionation column. When added to rPpChit1 or to midgut lysates, PpChit1 sera reduced chitinolytic activity by 65-70%.

Original languageEnglish (US)
Pages (from-to)703-712
Number of pages10
JournalInsect Molecular Biology
Volume14
Issue number6
DOIs
StatePublished - Dec 1 2005
Externally publishedYes

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Keywords

  • Chitinolytic activity
  • Leishmania
  • Peritrophic matrix
  • Sand fly

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Insect Science

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