Protein aggregated into bacterial inclusion bodies does not result in protection from proteolytic digestion

Xavier Carbonell, Antonio Villaverde

Research output: Contribution to journalArticleResearchpeer-review

13 Citations (Scopus)

Abstract

Proteolytic resistance, as conferred by protein aggregation into inclusion bodies, has not been explored in detail. We have investigated the eventual digestion of several closely-related proteins, namely six insertional and two fusion mutants of the homotrimeric bacteriophage P22 tailspike (TSP) protein. When over-produced in E. coli, all these polypeptides form inclusion bodies accompanied by only traces of soluble protein. The mutations introduced in TSP impaired its degradation and enhanced its half live up to ten-fold, without affecting protein solubility. This indicates that protein properties other than solubility, are the main determinants of susceptibility to proteolysis. In addition, the analysis of the degradation fragments strongly suggests that the aggregated TSP polypeptides undergo a site-limited proteolytic attack, and that their complete digestion occurs through an in situ cascade cleavage process.
Original languageEnglish
Pages (from-to)1939-1944
JournalBiotechnology Letters
Volume24
DOIs
Publication statusPublished - 1 Dec 2002

Keywords

  • Protein aggregation
  • Protein folding
  • Proteolytic cascade
  • Recombinant protein
  • Tailspike protein

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