Ferricytochrome c encapsulated in silica nanoparticles: structural stability and functional properties.
Using a modified sol-gel technique, we have succeeded in encapsulating ferric cytochrome c in silica nanoparticles obtained from hydrolysis and polycondensation of tetramethylorthosilicate. Particles dimensions have been determined with dynamic light scattering; this technique yields an hydrodynamic radius of about 100 nm, each nanoparticle containing about 10(2)-10(3) proteins. If stored in the cold at low ionic strength, nanoparticles are stable for more than one week, even if a slow radius increase with time is observed. CD measurements show that encapsulated proteins exhibit substantially increased stability against guanidinium hydrochloride induced denaturation. Reduction kinetics of encapsulated ferric cytochrome c by sodium dithionite, measured with standard stopped flow techniques, are slower by a factor of ten with respect to those measured in solution. Analogous experiments with myoglobin suggest that this slowing down is due to the diffusion time of dithionite within the silica matrix. Indeed, if a smaller ligand like CO is used, the intrinsic kinetic properties of encapsulated proteins are found to be unaltered even in the millisecond time range. The reported data show that our nanoparticles are extremely useful both for basic research, to study the stability and functions of encapsulated proteins, and for their potential biotechnological applications.
Fiandaca
Gianluca
G
Istituto Nazionale per la Fisica della Materia and Dipartimento di Scienze Fisiche ed Astronomiche, University of Palermo, via Archirafi 36, I-90123 Palermo, Italy.
Vitrano
Eugenio
E
Cupane
Antonio
A
D000818:Animals; D015374:Biosensing Techniques; D002248:Carbon Monoxide / Q000737:chemistry; D002942:Circular Dichroism; D045304:Cytochromes c / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D019791:Guanidine / Q000737:chemistry / Q000494:pharmacology; D006736:Horses; D006868:Hydrolysis; D007477:Ions; D007700:Kinetics; D008024:Ligands; D008027:Light; D009211:Myoglobin / Q000737:chemistry; D011487:Protein Conformation; D012542:Scattering, Radiation; D017640:Silicates / Q000737:chemistry; D012825:Silicon / Q000737:chemistry*; D013997:Time Factors; D014867:Water / Q000737:chemistry
D007477:Ions; D008024:Ligands; D009211:Myoglobin; D017640:Silicates; D014867:Water; D002248:Carbon Monoxide; D045304:Cytochromes c; D019791:Guanidine; D012825:Silicon