title: P450scc mutant nanostructuring for optimal assembly.
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 pi-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.
authors.affiliation: Department of Biophysical M&O Sciences and Technologies, University of Genova, Genova 16132, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Ghisellini
authors.forename: Cristina
authors.initials: C
authors.lastname: Paternolli
authors.forename: Mirco
authors.initials: M
authors.lastname: Antonini
authors.forename: Claudio
authors.initials: C
authors.lastname: Nicolini
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D003001:Cloning, Molecular; D003198:Computer Simulation; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D004563:Electrochemistry / Q000379:methods; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D024861:Genetic Enhancement / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D036103:Nanotechnology / Q000379:methods*; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D020539:Sequence Analysis, Protein / Q000379:methods*; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D003577:Cytochrome P-450 Enzyme System