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Pressure effects on protein flexibility monomeric proteins

P Cioni1, G B Strambini

  • 1C.N.R.--Istituto di Biofisica, Pisa, Italy.

Journal of Molecular Biology
|September 23, 1994
PubMed
Summary

Hydrostatic pressure subtly alters protein flexibility, affecting monomeric proteins like apoazurin and ribonuclease T1. This study reveals pressure-induced conformational changes, impacting protein structure and stability.

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Area of Science:

  • Biophysics
  • Protein dynamics
  • Hydrostatic pressure effects

Background:

  • Monomeric proteins exhibit conformational flexibility.
  • Hydrostatic pressure can induce predenaturational changes in protein structure.
  • Tryptophan phosphorescence decay kinetics are sensitive to protein microenvironment.

Purpose of the Study:

  • To investigate pressure-induced alterations in monomeric protein flexibility using tryptophan phosphorescence.
  • To characterize the conformational changes in apoazurin, ribonuclease T1, and phosphoglycerate kinase under hydrostatic pressure (< or = 3 kbar).
  • To elucidate the role of protein location and hydration in pressure-induced flexibility changes.

Main Methods:

  • Analysis of tryptophan phosphorescence decay kinetics.
  • Measurement of triplet lifetime (tau) and decay amplitudes.
  • Application of hydrostatic pressure up to 3 kbar.
  • Use of glycerol as a co-solvent to modulate pressure effects.

Main Results:

  • Pressure induced subtle conformational changes in all studied proteins.
  • Apoazurin showed increased core rigidity, while ribonuclease T1 and phosphoglycerate kinase exhibited increased surface flexibility.
  • Phosphoglycerate kinase underwent a partial unfolding-like transition, influenced by temperature and abolished by glycerol.
  • Glycerol attenuated pressure effects on apoazurin and reversed them for ribonuclease T1, while eliminating transitions in phosphoglycerate kinase.

Conclusions:

  • Hydrostatic pressure universally affects protein flexibility through subtle conformational changes.
  • Pressure's impact on flexibility depends on the protein's structure and the probe's location.
  • Opposing effects of pressure on protein flexibility are explained by reduced internal cavities and increased hydration.

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