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PMID: 15761028 Published · ppublish English Journal Article Review

Imaging molecular interactions in living cells.

Molecular endocrinology (Baltimore, Md.) ·Vol. 19 ·No. 7 ·2005-07-00 ·Pages 1675-86

Day RN, Schaufele F

Abstract

Hormones integrate the activities of their target cells through receptor-modulated cascades of protein interactions that ultimately lead to changes in cellular function. Understanding how the cell assembles these signaling protein complexes is critically important to unraveling disease processes, and to the design of therapeutic strategies. Recent advances in live-cell imaging technologies, combined with the use of genetically encoded fluorescent proteins, now allow the assembly of these signaling protein complexes to be tracked within the organized microenvironment of the living cell. Here, we review some of the recent developments in the application of imaging techniques to measure the dynamic behavior, colocalization, and spatial relationships between proteins in living cells. Where possible, we discuss the application of these different approaches in the context of hormone regulation of nuclear receptor localization, mobility, and interactions in different subcellular compartments. We discuss measurements that define the spatial relationships and dynamics between proteins in living cells including fluorescence colocalization, fluorescence recovery after photobleaching, fluorescence correlation spectroscopy, fluorescence resonance energy transfer microscopy, and fluorescence lifetime imaging microscopy. These live-cell imaging tools provide an important complement to biochemical and structural biology studies, extending the analysis of protein-protein interactions, protein conformational changes, and the behavior of signaling molecules to their natural environment within the intact cell.

MeSH Terms
Animals Cells/chemistry Hormones/physiology Humans Luminescent Proteins/analysis Microscopy, Fluorescence Proteins/analysis Receptors, Cytoplasmic and Nuclear/physiology Spectrometry, Fluorescence/methods
Chemicals
Hormones Luminescent Proteins Proteins Receptors, Cytoplasmic and Nuclear
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Day Richard N
Department of Medicine, P.O. Box 800578, University of Virginia Health Sciences Center, Charlottesville, VA 22908, USA. rnd2v@virginia.edu
Schaufele Fred
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Article Info
Journal
Molecular endocrinology (Baltimore, Md.)
Abbr.
Mol Endocrinol
ISSN
0888-8809
DOI
10.1210/me.2005-0028
Email
rnd2v@virginia.edu
Published
2005-07-00
Epub
2005-00-10
Pages
1675-86
Language
English
Region
United States
NLM ID
8801431
PMCID
PMC2900770
Subset
IM
Grants
NIDDK NIH HHS · R01 DK043701 · United States
NIDDK NIH HHS · R01 DK043701-11 · United States
NIDDK NIH HHS · R01 DK054345 · United States
NIDDK NIH HHS · R01 DK054345-06 · United States
External Links
PubMed source DOI: 10.1210/me.2005-0028 PMC Full Text View journal details
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