Green fluorescent protein for in situ synthesis of highly uniform Au nanoparticles and monitoring protein denaturation.
Purified recombinant green fluorescent protein (GFP) expressed in E. coli was used for single-step synthesis of gold nanoparticles (Au NPs) with extraordinary size specificity in aqueous medium. The fluorescence of GFP offered a probe for concomitant changes in the protein during the course of synthesis, in addition to the monitoring of the time-dependent formation of Au NPs by the surface plasmon resonance. Reaction of AuCl(4)(-) with the protein produced spherical Au NPs having diameters ranging from 5-70 nm. Remarkably, addition of 1.0x10(-5) M AgNO(3) in the medium produced uniform spherical Au NPs with particle diameter of 2.2+/-0.5 nm. Fluorescence spectroscopic measurements suggest that during synthesis of Au NPs in absence of AgNO(3), partial denaturation of the protein occurred resulting in the lowering of fluorescence intensity. On the other hand, when the NPs were synthesized in the presence of AgNO(3) complete denaturation of the protein with complete loss of fluorescence could be observed, which was further confirmed by native and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE). However, use of AgNO(3) only resulted neither in the formation of NPs nor had any significant effect on the fluorescence of GFP.
Sanpui
Pallab
P
Centre for Nanotechnology, Indian Institute of Technology Guwahati, Guwahati 781039, India.
Pandey
Shivendra B
SB
Ghosh
Siddhartha Sankar
SS
Chattopadhyay
Arun
A
D004591:Electrophoresis, Polyacrylamide Gel; D006046:Gold / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry*; D007700:Kinetics; D053768:Metal Nanoparticles / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D036103:Nanotechnology / Q000379:methods; D010316:Particle Size; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012835:Silver Nitrate / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D006046:Gold; D012835:Silver Nitrate