Title :
P450scc mutant nanostructuring for optimal assembly
Author :
Ghisellini, Paola ; Paternolli, Cristina ; Antonini, Mirco ; Nicolini, Claudio
Author_Institution :
Dept. of Biophys. M&O Sci. & Technol., Univ. of Genova, Italy
fDate :
6/1/2004 12:00:00 AM
Abstract :
Molecular modeling and protein engineering were synergically employed to improve the fabrication of cytochrome P450scc mutant nanostructures for biodevice assembly. The optimization of protein three-dimensional structure by molecular modeling was performed using two models: in vacuum and simulating the presence of a polar solvent. Calculations were performed on a model to predict a P450scc mutant which could improve the process of molecules´ immobilization onto solid supports. Engineerized cytochrome P450scc thin films were prepared and characterized by various biophysical techniques such as π-A isotherms, surface potential measurements, Brewster angle microscopy, UV-vis spectroscopy, circular dichroism, nanogravimetry, and electrochemical analysis. This paper takes into consideration biomolecules modified by protein engineering that represent a new and powerful approach for obtaining synthetic simpler artificial structures with new or improved properties (i.e., specificity, stability, sensitivity, etc.) useful for biosensors development.
Keywords :
bioelectric potentials; biological techniques; biosensors; circular dichroism; electrochemical analysis; enzymes; molecular biophysics; molecular configurations; nanotechnology; optimisation; physiological models; surface potential; ultraviolet spectra; visible spectra; /spl pi/-A isotherms; Brewster angle microscopy; UV-vis spectroscopy; biodevice assembly; biomolecules; biosensors; circular dichroism; cytochrome P450scc mutant nanostructures; cytochrome P450scc thin films; electrochemical analysis; molecular modeling; nanogravimetry; optimal assembly; optimization; polar solvent; protein engineering; protein three-dimensional structure; surface potential measurements; Assembly; Fabrication; Goniometers; Nanostructures; Power engineering and energy; Predictive models; Protein engineering; Solid modeling; Solvents; Transistors; Amino Acid Sequence; Amino Acid Substitution; Cloning, Molecular; Computer Simulation; Cytochrome P-450 Enzyme System; Electrochemistry; Enzyme Stability; Escherichia coli; Genetic Enhancement; Models, Molecular; Molecular Sequence Data; Mutagenesis, Site-Directed; Nanotechnology; Protein Conformation; Protein Engineering; Sequence Analysis, Protein; Structure-Activity Relationship; Temperature;
Journal_Title :
NanoBioscience, IEEE Transactions on
DOI :
10.1109/TNB.2004.828267