Refolding and reconstitution of human recombinant Bax inhibitor-1 into liposomes from inclusion bodies expressed in Escherichia coli

Protein Expr Purif. 2009 Jul;66(1):35-8. doi: 10.1016/j.pep.2009.01.005. Epub 2009 Jan 22.

Abstract

In this study, we report the simultaneous refolding and reconstitution of the recombinant Bax inhibitor-1 (BI-1) from inclusion bodies expressed in Escherichia coli. A functional assay showed that the resulting proteoliposomes responded to acidic conditions and triggered the release of entrapped Ca(2+) from liposomes. The secondary structure of the reconstituted BI-1 was also determined using circular dichroism, which revealed an increase of alpha-helix content and a decrease of random structure when exposed to acidic solutions. These conformational changes may be responsible for the proton ion-induced Ca(2+) release of BI-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis Regulatory Proteins / chemistry
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism*
  • Calcium / metabolism
  • Escherichia coli / cytology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Inclusion Bodies / chemistry*
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Protein Engineering
  • Protein Folding
  • Protein Structure, Secondary
  • Proteolipids / chemistry*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*

Substances

  • Apoptosis Regulatory Proteins
  • Membrane Proteins
  • Proteolipids
  • Recombinant Fusion Proteins
  • TMBIM6 protein, human
  • proteoliposomes
  • Calcium