Spectroscopic investigation of structure in octarellin (a de novo protein designed to adopt the alpha/beta-barrel packing).
We present here a spectroscopic structural characterization of octarellin, a recently reported de novo protein modelled on alpha/beta-barrel proteins [K. Goraj, A. Renard and J.A. Martial (1990) Protein Engng, 3, 259-266]. Infrared and Raman spectra analyses of octarellin's secondary structure reveal the expected percentage of alpha-helices (30%) and a higher beta-sheet content (40%) than predicted from the design. When the Raman spectra obtained with octarellin and native triosephosphate isomerase (a natural alpha/beta-barrel) are compared, similar percentages of secondary structures are found. Thermal denaturation of octarellin monitored by CD confirms that its secondary structures are quite stable, whereas its native-like tertiary fold is not. Tyrosine residues, predicted to be partially hidden from solvent, are actually exposed as revealed by Raman and UV absorption spectra. We conclude that the attempted alpha/beta-barrel conformation in octarellin may be loosely packed. The criteria used to design octarellin are discussed and improvements suggested.
Beauregard
M
M
Laboratorie de Biologie Moléculaire et de Génie Génétique, Université de Liège, Institut de Chimie, Sart Tilman, Belgium.
Goraj
K
K
Goffin
V
V
Heremans
K
K
Goormaghtigh
E
E
Ruysschaert
J M
JM
Martial
J A
JA
D002942:Circular Dichroism; D005583:Fourier Analysis; D011487:Protein Conformation; D011489:Protein Denaturation; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry*; D013055:Spectrophotometry, Infrared; D013057:Spectrum Analysis; D013059:Spectrum Analysis, Raman
D011994:Recombinant Proteins; C073399:octarellin