Labeling ribonuclease S with a 3 nm Au nanoparticle by two-step assembly.
We label ribonuclease S with a 3 nm Au nanoparticle (NP) by utilizing its two-piece structure. One portion, S-peptide, is mutated with a unique NP attachment site. NP-peptide self-assembles with the other portion, S-protein, to form an active enzyme. NP mobility decreases with peptide labeling and S-protein association. Surface plasmon shifts support conjugation. Higher S-peptide coverages on the NP surface reduce nonspecific adsorption, while sterically hindering assembly of RNaseS. Thiols displace nonspecific adsorption, maximizing site-specific labeling.
Aubin
Marie-Eve
ME
Department of Mechanical Engineering and Biological Engineering Division, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
Morales
Diana G
DG
Hamad-Schifferli
Kimberly
K
D003460:Crystallization / Q000379:methods*; D008027:Light; D008422:Materials Testing; D036103:Nanotechnology / Q000379:methods; D043942:Nanotubes / Q000032:analysis / Q000737:chemistry* / Q000648:ultrastructure*; D010316:Particle Size; D011485:Protein Binding; D012260:Ribonucleases / Q000032:analysis / Q000737:chemistry* / Q000648:ultrastructure*; D012834:Silver / Q000032:analysis / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods; D020349:Surface Plasmon Resonance / Q000379:methods*
D012834:Silver; D012260:Ribonucleases; C022542:ribonuclease S