Expression of mRNAs coding for VAP1/crotastatin-like metalloproteases in the venom glands of three South American pit vipers assessed by quantitative real-time PCR

Toxicon. 2008 Dec 15;52(8):897-907. doi: 10.1016/j.toxicon.2008.08.022. Epub 2008 Sep 27.

Abstract

Snake venom metalloproteases encompass a large family of toxins, with approximately 200 members already catalogued, which exhibit a diversity of structures and biological functions. From this relatively large number, only a dozen examples of apoptosis-inducing metalloproteases, like VAP1 and 2 from the venom of Crotalus atrox, are known. Since most VAP1-like toxins ever characterized were purified from the venom of Viperidae species inhabiting diverse places on earth, we investigate the expression of VAP-like metalloproteases in the venom gland of three representative pit vipers of the Brazilian territory. By molecular cloning and quantitative real-time polymerase chain reaction, using as calibrator gene the Crotalus durissus terrificus homolog of VAP1, named crotastatin, it is reported here that VAP1/crotastatin-like homologues in the venom gland of Bothrops atrox, C. d. cascavella and Lachesis m. rhombeata are expressed at different levels. Hence, batroxstatins, the crotastatin-like precursors from B. atrox, are expressed 87 times more than crotastatin-1, from C. d. cascavella, and 7.5-fold that lachestatins, from L. m. rhombeata. Moreover, in silico structural analysis of amino acid sequences indicates that batroxstatin-2, crotastatins and lachestatin-1 and -2 which share the archetypal motifs and metal- binding sites of VAP1, are subgrouped in a branch that comprises some apoptosis-inducing toxins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Apoptosis Regulatory Proteins / genetics*
  • Apoptosis Regulatory Proteins / metabolism
  • Base Sequence
  • Computer Simulation
  • Crotalid Venoms / chemistry
  • Crotalid Venoms / genetics*
  • Crotalid Venoms / metabolism
  • Crotalus / genetics*
  • Crotalus / metabolism
  • Gene Expression
  • Gene Library
  • Linear Models
  • Metalloendopeptidases / genetics*
  • Metalloendopeptidases / metabolism
  • Metalloproteases / genetics
  • Metalloproteases / metabolism
  • Molecular Sequence Data
  • Phylogeny
  • Polymerase Chain Reaction
  • Polymorphism, Genetic
  • RNA, Messenger / analysis
  • Sequence Alignment

Substances

  • Apoptosis Regulatory Proteins
  • Crotalid Venoms
  • RNA, Messenger
  • Metalloproteases
  • Metalloendopeptidases
  • vascular apoptosis-inducing protein 1, Crotalus atrox