Structure of lysozyme dissolved in neat organic solvents as assessed by NMR and CD spectroscopies.
The structure of the model protein hen egg-white lysozyme dissolved in water and in five neat organic solvents (ethylene glycol, methanol, dimethylsulfoxide (DMSO), formamide, and dimethylformamide (DMF)) has been examined by means of 1H NMR and circular dichroism (CD) spectroscopies. The NMR spectra of lysozyme reveal the lack of a defined tertiary structure in all five organic solvents, although the examination of line widths suggests the possibility of some ordered structure in ethylene glycol and in methanol. The near-UV CD spectra of the protein suggest no tertiary structure in lysozyme dissolved in DMSO, formamide, and DMF, while a distinctive (albeit less pronounced than in water) tertiary structure is seen in ethylene glycol and a drastically changed one in methanol. A highly developed secondary structure was observed by far-UV CD in ethylene glycol and methanol; interestingly, the alpha-helix content of the protein in both was greater than in water, while the beta-structure content was lower. (Solvent absorbance in the far-UV region prevents conclusions about the secondary structure in DMSO, formamide and DMF.)
Knubovets
T
T
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Osterhout
J J
JJ
Klibanov
A M
AM
D000818:Animals; D002645:Chickens; D002942:Circular Dichroism; D004121:Dimethyl Sulfoxide; D004126:Dimethylformamide; D019855:Ethylene Glycol; D005559:Formamides; D000432:Methanol; D009113:Muramidase / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D011487:Protein Conformation; D012996:Solutions; D012997:Solvents
D005559:Formamides; D012996:Solutions; D012997:Solvents; C031066:formamide; D004126:Dimethylformamide; D009113:Muramidase; D019855:Ethylene Glycol; D000432:Methanol; D004121:Dimethyl Sulfoxide