Tracing the dynamic distribution of endogenous fluorescent protein in tumor cells by FRAP

Ying Jin; Da Xing; Yonghong Tang

doi:10.1117/12.572444

29 July 2004 Tracing the dynamic distribution of endogenous fluorescent protein in tumor cells by FRAP

Ying Jin, Da Xing, Yonghong Tang

Proceedings Volume 5486, ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics; (2004) https://doi.org/10.1117/12.572444
Event: ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics, 2003, Silsoe, United Kingdom

Abstract

The NPC is the portal for the exchange of proteins, mRNA, and ions between nucleus and cytoplasm. Many small molecules (<10 kDa) permeate the nucleus by simple diffusion through the pore, but molecules larger than 70 kDa require ATP and a nuclear localization sequence for their transport. In isolated Xenopus oocyte nuclei, diffusion of intermediate-sized molecules appears to be regulated by the NPC, dependent upon Ca²⁺ in the nuclear envelope. We have applied real-time imaging and fluorescence recovery after photobleaching to examine the nuclear pore permeability of 27-kDa EGFP in single intact cells. We found that EGFP diffused bidirectionally via the NPC across the nuclear envelope. Although diffusion is slowed several-decade-fold at the nuclear envelope boundary compared to diffusion within the nucleus or cytoplasm, this delay is expected for the reduced cross-sectional area of the NPCs.

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Ying Jin, Da Xing, and Yonghong Tang "Tracing the dynamic distribution of endogenous fluorescent protein in tumor cells by FRAP", Proc. SPIE 5486, ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics, (29 July 2004); https://doi.org/10.1117/12.572444

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KEYWORDS

Luminescence

Diffusion

Molecules

Proteins

Confocal microscopy

Fluorescent proteins

Green fluorescent protein

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