TY - CHAP
T1 - Visualization of protein interactions in living cells
AU - Zal, Tomasz
PY - 2008
Y1 - 2008
N2 - Ligand binding to cell membrane receptors sets off a series of protein interactions that convey the nuances of ligand identity to the cell interior. The information may be encoded in conformational changes, the interaction kinetics and, in the case of multichain immunoreceptors, by chain rearrangements. The signals may be modulated by dynamic compartmentalization of the cell membrane, cellular architecture, motility, and activation-all of which are difficult to reconstitute for studies of receptor signaling in vitro. In this chapter, we will discuss how protein interactions in general and receptor signaling in particular can be studied in living cells by different fluorescence imaging techniques. Particularly versatile are methods that exploit Förster resonance energy transfer (FRET), which is exquisitely sensitive to the nanometer-range proximity and orientation between fluorophores. Fluorescence correlation microscopy (FCM) can provide complementary information about the stoichiometry and diffusion kinetics of large complexes, while bimolecular fluorescence complementation (BiFC) and other complementation techniques can capture transient interactions. A continuing challenge is extracting from the imaging data the quantitative information that is necessary to verify different models of signal transduction.
AB - Ligand binding to cell membrane receptors sets off a series of protein interactions that convey the nuances of ligand identity to the cell interior. The information may be encoded in conformational changes, the interaction kinetics and, in the case of multichain immunoreceptors, by chain rearrangements. The signals may be modulated by dynamic compartmentalization of the cell membrane, cellular architecture, motility, and activation-all of which are difficult to reconstitute for studies of receptor signaling in vitro. In this chapter, we will discuss how protein interactions in general and receptor signaling in particular can be studied in living cells by different fluorescence imaging techniques. Particularly versatile are methods that exploit Förster resonance energy transfer (FRET), which is exquisitely sensitive to the nanometer-range proximity and orientation between fluorophores. Fluorescence correlation microscopy (FCM) can provide complementary information about the stoichiometry and diffusion kinetics of large complexes, while bimolecular fluorescence complementation (BiFC) and other complementation techniques can capture transient interactions. A continuing challenge is extracting from the imaging data the quantitative information that is necessary to verify different models of signal transduction.
UR - http://www.scopus.com/inward/record.url?scp=58149340844&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=58149340844&partnerID=8YFLogxK
U2 - 10.1007/978-0-387-09789-3_14
DO - 10.1007/978-0-387-09789-3_14
M3 - Chapter
C2 - 19065792
AN - SCOPUS:58149340844
SN - 9780387097886
T3 - Advances in Experimental Medicine and Biology
SP - 183
EP - 197
BT - Multichain Immune Recognition Receptor Signaling
A2 - Sigalov, Alexander
ER -