Purification and Characterization of Crystalins by Aqueous Two-Phase Extraction

  • Bermudez, Ondrea (Chemical Engineering Department, University of Missouri-Rolla, Rolla, MO65409, USA) ;
  • Forciniti, Daniel (Chemical Engineering Department, University of Missouri-Rolla, Rolla, MO65409, USA)
  • 발행 : 2001.12.01

초록

Crystallins are a family of water-soluble proteins that constitute up to 90% of the wa-ter-soluble proteins in mammalian eye lenses, We present in this paper an alternative purification method for these proteins using polyethylene glycol/dextran aqueous two-phase extraction. Un-der the appropriate conditions, we were able to recover the γ-crystallin fraction essentially free of the remaining proteins. High concentrations of salt at a neutral pH maximize the recovery of γ-crystallins in the top phase and minimize the contamination by the other proteins present in the lenses. The proposed protocol decreases the separation time by about 50%. The complex partition behavior observed for these proteins reflects a delicate balance between protein/phase-forming species(various polymers and salts) and protein interactions. This is evidenced, in part, by the role played by the largest proteins in this group as a "pseudo"phase-forming species.

키워드

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