S-Space College of Medicine/School of Medicine (의과대학/대학원) Nuclear Medicine (핵의학전공) Journal Papers (저널논문_핵의학전공)
Bioimaging of geographically adjacent proteins in a single cell by quantum dot-based fluorescent resonance energy transfer
- Kang, Won Jun; Ko, Mee Hyang; Lee, Dong Soo; Kim, Soonhag
- Issue Date
- WILEY-V C H VERLAG GMBH
- PROTEOMICS CLINICAL APPLICATIONS; Vol.3 12; 1383-1388
- Thousands of proteins are simultaneously involved in the maintenance of a single cancer cell. Fluorescent resonance energy transfer (FRET) is one of the most general techniques for imaging biologically interacting molecules in a cell. Here, we applied FRET to image the colocalization of two proteins that do not interact biologically (nucleolin and integrin alpha(v)beta(3)), both of which are highly expressed in the plasma membrane of cancer cells. AS1411 aptamer, which targets nucleolin, was labeled by Cy3 (Cy3-AS1411) and arginine-glycine-aspartic acid (RGD) peptide, which targets integrin alpha(v)beta(3), was conjugated with quantum dot (525 nm, Qd) Qd arginine-glycine-aspartic acid (Qd-RGD). FRET activities between Cy3-AS1411 and Qd-RGD were measured in HeLa cells, a human cervical cancer cell line. FRET phenomena between Qd and Cy3 showed good compatibility according to proximity. The fluorescence signature using Qd-RGD and Cy3-AS1411 showed that nucleolin and integrin alpha(v)beta(3) proteins were highly expressed in HeLa cells. Co-incubation of Qd-RGD and Cy3-AS1411 in a single HeLa cell demonstrated that the fluorescence overlay by FRET was quantitatively and geographically quite different from that of individual confocal images. These results suggest that Qd-based FRET analysis can provide information on geographical co-localization of proteins in naive cells, which is very important for determining the molecular and cellular functions of genes involved in cancers and other clinical diseases.
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