title: Alcohol-induced molten globule intermediates of proteins: are they real folding intermediates or off pathway products?
abstract: Alcohols have been shown to cause a conformational transition of proteins into a new stable conformational state resembling that of the "molten globule intermediate" characterized by high alpha-helical content and disrupted tertiary structure. We have studied the effect of monohydric alcohols on the stability and structural characteristics of small globular protein hen egg white lysozyme by the combined use of differential scanning calorimetry, circular dichroism, and nuclear magnetic resonance spectroscopy. The protein stability was found to be significantly decreased with increasing alcohol concentration, and, in presence of moderate to higher alcohol concentrations, depending on the pH and alcohol studied, the protein was found to be unfolded even at 4 degrees C. Correlation between thermal stability and alpha-helicity of several small globular proteins like hen egg white lysozyme, horse heart cytochrome C, and bovine carbonic anhydrase B, observed in presence of increasing alcohol concentrations, suggests that probably alcohols induce helical structures in unfolded protein. The temperature-dependent near- and far-UV circular dichroism and proton nuclear magnetic resonance spectroscopic studies on lysozyme in the presence of 2,2,2-trifluoroethanol and methanol, respectively, showed that alcohols do induce significantly higher helical structures in unfolded protein compared to folded protein. The results presented in this paper suggest that the molten globule intermediate of proteins in the presence of high alcohols as reported earlier is due to alcohol-induced local folding rather than global folding of unfolded protein and hence is an off-pathway product and not a real folding intermediate.
authors.affiliation: Division of Membrane Biology, Central Drug Research Institute, Lucknow, India. root@cscdri.ren.nic.in
authors.forename: V
authors.initials: V
authors.lastname: Bhakuni
mesh_terms: D000438:Alcohols / Q000737:chemistry* / Q000494:pharmacology; D002152:Calorimetry, Differential Scanning; D002256:Carbonic Anhydrases / Q000737:chemistry; D003574:Cytochrome c Group / Q000737:chemistry / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004795:Enzyme Stability / Q000187:drug effects; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol / Q000494:pharmacology; D009113:Muramidase / Q000737:chemistry / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D017433:Protein Structure, Secondary / Q000187:drug effects; D011506:Proteins / Q000737:chemistry* / Q000187:drug effects; D013696:Temperature; D013816:Thermodynamics; D014270:Trifluoroethanol / Q000494:pharmacology
chemical_list: D000438:Alcohols; D003574:Cytochrome c Group; D011506:Proteins; D014270:Trifluoroethanol; D009113:Muramidase; D002256:Carbonic Anhydrases; D000432:Methanol