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.

Determination of Protein-Protein Interaction for Cancer Control via Mass Spectrometry and Nanoconductimetry of NAPPA SNAP Arrays: An Overview

“…The P450scc-cholesterol interaction has been well characterized in the extant literature and the results obtained have been satisfactorily compared with those in literature [28].…”

Matrices for Sensors from Inorganic, Organic, and Biological Nanocomposites

“…Cytochrome P450scc was found to be suitable for nanostructure assembly by LB technique combined with site directed mutagenesis [103,104,105]. Molecular modeling and protein engineering were synergically employed to improve the fabrication of cytochrome P450scc mutant nanostructures for biodevice assembly.…”

“…Above: NAPPA technology [103]. Below left: NAPPA region image with human kinase proteins acquired by the DNASER device.…”

Site‐directed mutations (Asp405Ile and Glu124Ile) in cytochrome P450scc: Effect on adrenodoxin binding

“…The expression plasmid pTrc99A-P450scc was obtained as described in Ghisellini et al [2004]. Escherichia coli strains JM109 cells were obtained from Stragene (La Jolla, CA).…”

“…To do that, it was used the program Discover (Molecular Simulation, Inc.). A comparison between mutants and the optimized cytochrome P450scc structure described in Ghisellini et al [2004] was performed.…”