Isolation and identification of insect intestinal mucin HaIIM86--the new target for Helicoverpa armigera biocontrol

Int J Biol Sci. 2011 Mar 19;7(3):286-96. doi: 10.7150/ijbs.7.286.

Abstract

There are many more glycoproteins in Helicoverpa armigera peritrophic membrane than midgut separated by SDS-PAGE analysis after Periodic acid-Schiff (PAS) and coomassie staining. The peritrophic membrane (PM) of H. armigera larvae contains about forty associated proteins. A cDNA library was constructed from H. armigera midgut mRNA to study the new target for pest biocontrol. An antiserum against Spodoptera exigua integral/total PM proteins cross reacted with several H. armigera PM proteins and was used to isolate a complete cDNA encoding an insect intestinal mucin (HaIIM86). The deduced protein sequence of the cDNA contained one potentially glycosylated, mucin-like domain, five cysteine-rich chitin-binding domains (CBDs) and two D-G rich regions. Mucin domain was lined between the first and second CBDs; the two additional D-G rich regions were proposed to internal reside at the amino terminus of the protein flanked by three cysteine-rich CBDs. HaIIM86 contains two D-G-rich tandem repeat domains flanked by cysteine-rich sequences in peritrophic membrane proteins which is not present in all the currently known PM proteins. Howerer the functions of D-G rich domains require further investigation. HaIIM86 was shown as 200 kDa protein by SDS-PAGE analysis and appeared to be associated with the PM. HaIIM86 has chitin-binding activity and can be degraded into 90 and 70 kDa by HaGV Enhancin in vivo. The finding has shown that HaIIM86 is the target substrate for enhancin and the potential target for pest control.

Keywords: Helicoverpa armigera; Peritrophic matrix; biocontrol; cDNA expression library; insect intestinal mucin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Chitin / metabolism
  • Cloning, Molecular
  • Gene Library
  • Insect Proteins / chemistry
  • Insect Proteins / genetics*
  • Insect Proteins / physiology
  • Larva / metabolism
  • Molecular Sequence Data
  • Moths / genetics*
  • Moths / growth & development
  • Moths / metabolism
  • Mucins / chemistry
  • Mucins / genetics*
  • Mucins / physiology
  • Pest Control, Biological
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Sequence Analysis, Protein

Substances

  • Insect Proteins
  • Mucins
  • Chitin