Intracellular imaging of targeted proteins labeled with quantum dots.
We developed a new method for imaging the movement of targeted proteins in living cancer cells with photostable and bright quantum dots (QDs). QDs were conjugated with various molecules and proteins, such as phalloidin, anti-tubulin antibody and kinesin. These bioconjugated QDs were mixed with a transfection reagent and successfully internalized into living cells. The movements of individual QDs were tracked for long periods of time. Phalloidin conjugated QDs bound to actin filaments and showed almost no movement. In contrast, anti-tubulin antibody conjugated QDs bound to microtubules and revealed dynamic movement of microtubules. Kinesin showed an interesting behavior whereby kinesin came to be almost paused briefly for a few seconds and then moved once again. This is in direct contrast to the smoothly continuous movement of kinesin in an in vitro assay. The maximum velocity of kinesin in cells was faster than that in the in vitro assay. These results suggest that intracellular movement of kinesin is different from that in the in vitro assay. This newly described method will be a powerful tool for investigating the functions of proteins in living cells.
Department of Material Science, Graduate School of Engineering, Tohoku University, Sendai, Japan.
Jungwoo
J
Yoo
Taketoshi
T
Kambara
Kohsuke
K
Gonda
Hideo
H
Higuchi
D021621:Imaging, Three-Dimensional / Q000379:methods; D016547:Kinesins / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010590:Phalloidine / Q000378:metabolism; D021381:Protein Transport; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D045663:Quantum Dots
D011506:Proteins; D010590:Phalloidine; D016547:Kinesins