Conjugation of active iron superoxide dismutase to nanopatterned surfaces.
Superoxide dismutase enzymes (SODs) are an essential part of the first line of cellular defense system against free radicals species. They catalyze the dismutation of superoxide radicals into oxygen and hydrogen peroxide. Although several studies have examined the attachment of superoxide dismutases to nanoparticles and nanostructures, never has been used a member of the Fe/MnSOD family. In this study, the behavior of plant origin FeSOD enzyme on three different nanopatterned surfaces was investigated as a function of covalent and electrostatic binding. Fluorescence microscopy was used to demonstrate that the protein is attached only to the gold layer. We also examined the activity of SOD by a colorimetric assay, and we have shown that the enzyme remains active after attachment to the three different surfaces under both kind of binding (electrostatic and covalent). This methodology could be useful for those who want to functionalize nanostructures with a SOD enzyme and test the activity. This process could be of great interest for the development of peroxynitrite and superoxide biosensors.
FideNa (Foundation for r+d in nanotechnology), Centro Jerónimo de Ayanz, C/Tajonar s/n, E-31006 Pamplona, Navarra, Spain.
Edurne
E
Tellechea
Iñaki
I
Cornago
Paula
P
Ciaurriz
Jose F
JF
Moran
Aaron C
AC
Asensio
D001391:Azo Compounds / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D007887:Fabaceae / Q000201:enzymology; D006046:Gold / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry; D008856:Microscopy, Fluorescence; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D012822:Silicon Dioxide / Q000737:chemistry; D055672:Static Electricity; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000378:metabolism; D013499:Surface Properties
D001391:Azo Compounds; D004800:Enzymes, Immobilized; D010940:Plant Proteins; C033302:diformazan dye; D006046:Gold; D012822:Silicon Dioxide; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase