Visualizing the self-assembly of tubulin with luminescent nanorods.
We succeeded in labeling tubulin with luminescent CdSe nanorods. The labeled tubulin was still able to self-assemble into microtubules, demonstrating that the protein has remained functional. The comparison with rhodamine-labeled tubulin revealed that whereas the rhodamine bleached completely within the observed time frame, the nanorods did not show any bleaching, which makes it possible to follow the key events of self-assembly at high time resolution. Preparation and derivatization of nanorods are described. Water-soluble, protein-reactive group-containing particles were covalently coupled to tubulin. The conjugate was purified by one cycle of assembly and disassembly and used to induce the formation of fluorescent microtubules. These results suggest that the use of luminescent nanorods should allow continuous confocal monitoring of dynamic biological processes.
Riegler
Jürgen
J
Freiburg Materials Research Center, D-79104 Freiburg, Germany.
Nick
Peter
P
Kielmann
Udo
U
Nann
Thomas
T
D001672:Biocompatible Materials / Q000138:chemical synthesis / Q000737:chemistry; D019187:Cadmium Compounds / Q000138:chemical synthesis / Q000737:chemistry*; D019281:Dimerization; D005456:Fluorescent Dyes; D006160:Guanosine Triphosphate / Q000737:chemistry; D049449:Luminescence; D008856:Microscopy, Fluorescence / Q000379:methods*; D008870:Microtubules / Q000737:chemistry / Q000648:ultrastructure*; D036103:Nanotechnology / Q000379:methods*; D043942:Nanotubes; D011485:Protein Binding; D011487:Protein Conformation; D045663:Quantum Dots; D013194:Staining and Labeling / Q000379:methods*; D013440:Sulfides / Q000138:chemical synthesis / Q000737:chemistry*; D014404:Tubulin / Q000138:chemical synthesis / Q000737:chemistry / Q000648:ultrastructure*
D001672:Biocompatible Materials; D019187:Cadmium Compounds; D005456:Fluorescent Dyes; D013440:Sulfides; D014404:Tubulin; C034939:cadmium sulfide; D006160:Guanosine Triphosphate