Influence of cyclodextrin ring substituents on folding-related aggregation of bovine carbonic anhydrase

Loretta Sharma, Ajit Sharma

Research output: Contribution to journalArticlepeer-review

60 Scopus citations

Abstract

A common obstacle to proper renaturation of an unfolded protein is aggregation, an intermolecular side reaction of immense importance in biotechnology and in the pathogenesis of several neurodegenerative diseases. Cyclic sugars known as cyclodextrins have been used as protein-folding aids. The effect of cyclodextrin chemistry on aggregation and refolding of carbonic anhydrase was evaluated in this study. Size-exclusion HPLC showed that cyclodextrins inhibit aggregate formation without interfering with the correct renaturation of carbonic anhydrase. PAGE of refolded enzyme provides further evidence of inhibition of folding-related aggregation by natural and chemically modified cyclodextrins. Although the amount of aggregate formed and recovery of active enzyme was dependent on cavity size, the nature of the chemical substituents found on the rims of the sugar molecule seems to play a more important role in cyclodextrin-assisted refolding of carbonic anhydrase. In general, neutral or cationic cyclodextrins with small cavities were found to be better folding aids than anionic cyclodextrins with larger holes. Although the exact prediction of the effect of a cyclodextrin substitution on protein refolding is not possible at present, these results clearly show that modified cyclodextrins can be designed that effectively inhibit protein aggregation.

Original languageEnglish
Pages (from-to)2456-2463
Number of pages8
JournalEuropean Journal of Biochemistry
Volume268
Issue number8
DOIs
StatePublished - 2001

Keywords

  • Aggregation
  • Carbonic anhydrase
  • Cyclodextrin
  • Refolding
  • Renaturation

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