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The protective effects of osmolytes on yeast alcohol dehydrogenase conformational stability and aggregation

Hong-Yan Han1, Zhi-Gang Yao, Cheng-Liang Gong

  • 1Department of Biology, Medical College of Soochow University, Jiangsu, Suzhou 215123, PR China.

Protein and Peptide Letters
|April 29, 2010
PubMed

Insights

Four osmolytes protect yeast alcohol dehydrogenase (YADH) from denaturation and aggregation. These compounds enhance protein stability and prevent clumping during refolding, offering insights into protein protection mechanisms.

Area of Science:

  • Biochemistry
  • Protein Chemistry
  • Enzymology

Background:

  • Yeast alcohol dehydrogenase (YADH) is a crucial enzyme.
  • Protein denaturation and aggregation can lead to loss of function.
  • Osmolytes are known to stabilize proteins.

Purpose of the Study:

  • To investigate the protective effects of four osmolytes on YADH.
  • To assess the impact of osmolytes on YADH's conformational stability and aggregation.
  • To understand the mechanism of osmolyte-mediated protein protection.

Main Methods:

  • Denaturation of YADH using guanidine.
  • Addition of four osmolytes: trehalose, dimethyl sulfoxide, glycine, and proline.
  • Measurement of inactivation rate constants (ki).
  • Determination of transition free energy changes (ΔΔGi) at 25°C.
  • Measurement of denaturation midpoint values (Cm).
  • Assessment of ANS-binding fluorescence intensity.
  • Observation of YADH aggregation during refolding.

Main Results:

  • The four osmolytes significantly protected YADH against guanidine-induced unfolding and inactivation.
  • Osmolytes reduced inactivation rate constants (ki).
  • Osmolytes increased transition free energy changes (ΔΔGi) and denaturation midpoint values (Cm).
  • Osmolytes decreased ANS-binding fluorescence intensity, indicating reduced exposure of hydrophobic regions.
  • Osmolytes prevented YADH aggregation in a concentration-dependent manner during refolding.

Conclusions:

  • Trehalose, dimethyl sulfoxide, glycine, and proline effectively stabilize YADH conformation.
  • These osmolytes protect YADH by inhibiting unfolding and aggregation.
  • The findings provide valuable insights into osmolyte-based strategies for protein stabilization.

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