Synchrotron radiation circular dichroism spectroscopy applied to metmyoglobin and a 4-alpha-helix bundle carboprotein

Biopolymers. 2005 May;78(1):46-52. doi: 10.1002/bip.20253.

Abstract

The novel technique, synchrotron radiation-based circular dichroism (SR-CD), has been applied to the study of metmyoglobin and a carboprotein (carbohydrate-based peptide with protein tertiary structure) with 4-alpha-helix bundle structure, as well as a carbopeptide (carbohydrate-based peptide) with a truncated peptide sequence. The use of synchroton radiation (SR) enabled circular dichroism (CD) measurements in the vacuum ultraviolet (VUV) down to 168 nm in D(2)O and 160 nm in 2,2,2-trifluoroethanol (TFE). The band shape in the CD spectra in the low wavelength region was studied, comparing samples with two types of alpha-helical tertiary structure, namely the globin fold and the 4-alpha-helix bundle motif. No significant differences were found between the CD spectra of the alpha-helical samples (metmyoglobin and carboprotein) in D(2)O solution. The use of 2,2,2-TFE (TFE) as solvent clearly alters the VUV CD but the two samples have very similar CD spectra. The solvent-induced denaturing of metmyoglobin in TFE was observed using absorption and CD spectroscopy of the Soret band, with results indicating heme release. The VUV spectrum of TFE-denatured metmyoglobin exhibits dramatic differences in comparison with previous studies of the native enzyme in aqueous solution. The implications of this observation are discussed.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Circular Dichroism*
  • Galactose / analogs & derivatives*
  • Galactose / chemistry
  • Horses
  • Hydrogen Bonding
  • Hydrogen-Ion Concentration
  • Metmyoglobin / chemistry*
  • Metmyoglobin / drug effects
  • Peptides / chemical synthesis
  • Peptides / chemistry*
  • Protein Denaturation
  • Protein Folding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Solvents
  • Spectrophotometry, Ultraviolet
  • Synchrotrons*
  • Trifluoroethanol / pharmacology
  • Water / chemistry

Substances

  • Peptides
  • Solvents
  • methyl-galactopyranoside
  • Water
  • Metmyoglobin
  • Trifluoroethanol
  • Galactose