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Disulfide linkages in Plasmodium falciparum plasmepsin-i are essential elements for its processing activity and

Sirisak Lolupiman1, Pilaiwan Siripurkpong2, Jirundon Yuvaniyama1

  • 1Department of Biochemistry and Center for Excellence in Protein Structure and Function, Faculty of Science, Mahidol University, Bangkok, Thailand.

Plos One
|March 4, 2014
PubMed

Insights

Researchers improved the yield of soluble Plasmodium falciparum plasmepsin-I (PM-I), a malaria drug target, by optimizing protein refolding and disulfide bond formation. This strategy enables better structural studies for rational drug design against malaria.

Area of Science:

  • Biochemistry
  • Parasitology
  • Structural Biology

Background:

  • Plasmodium falciparum plasmepsin-I (PM-I) is a potential drug target for treating human malaria.
  • Obtaining soluble PM-I for structural studies is challenging due to difficulties in heterologous expression and low yields.

Purpose of the Study:

  • To develop a strategy for improved yield of soluble, active PM-I.
  • To facilitate structural determination of PM-I for rational inhibitor design.

Main Methods:

  • Optimized heterologous expression of semi-pro-PM-I in E. coli C41(DE3).
  • Investigated protein refolding conditions, focusing on disulfide bond formation.
  • Assessed autoprocessing and activation of refolded PM-I.

Main Results:

  • Successfully excluded truncated semi-pro-PM-I expression.
  • Identified structural instability in acidic conditions due to incomplete disulfide bonds as a cause of low yield.
  • Achieved significantly improved yields of 10 mg/L semi-pro-PM-I, yielding 6-8 mg/L mature PM-I.

Conclusions:

  • Optimized refolding and disulfide bond formation strategies enhance soluble PM-I yield.
  • This improved yield facilitates structural studies and drug development for malaria.
  • The strategy overcomes previous limitations in obtaining sufficient quantities of active PM-I.

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