Visualization of Protein Interactions in Living Cells

Overview

Journal Adv Exp Med Biol

Specialties Biology
General Medicine

Date 2008 Dec 11

PMID 19065792

Citations 6

Authors

Tomasz Zal

Affiliations

Soon will be listed here.

Abstract

Ligand binding to cell membrane receptors sets off a series of protein interactions that convey the nuances ofligand 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.

Citing Articles

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References

Muller B, Zaychikov E, Brauchle C, Lamb D . Pulsed interleaved excitation. Biophys J. 2005; 89(5):3508-22. PMC: 1366845. DOI: 10.1529/biophysj.105.064766. View

Young R, Arnette J, Roess D, Barisas B . Quantitation of fluorescence energy transfer between cell surface proteins via fluorescence donor photobleaching kinetics. Biophys J. 1994; 67(2):881-8. PMC: 1225431. DOI: 10.1016/S0006-3495(94)80549-1. View

Peter M, Ameer-Beg S, Hughes M, Keppler M, Prag S, Marsh M . Multiphoton-FLIM quantification of the EGFP-mRFP1 FRET pair for localization of membrane receptor-kinase interactions. Biophys J. 2004; 88(2):1224-37. PMC: 1305125. DOI: 10.1529/biophysj.104.050153. View

Zal T, Zal M, Gascoigne N . Inhibition of T cell receptor-coreceptor interactions by antagonist ligands visualized by live FRET imaging of the T-hybridoma immunological synapse. Immunity. 2002; 16(4):521-34. DOI: 10.1016/s1074-7613(02)00301-1. View

Mittler R, Goldman S, Spitalny G, Burakoff S . T-cell receptor-CD4 physical association in a murine T-cell hybridoma: induction by antigen receptor ligation. Proc Natl Acad Sci U S A. 1989; 86(21):8531-5. PMC: 298316. DOI: 10.1073/pnas.86.21.8531. View

Tolar P, Sohn H, Pierce S . The initiation of antigen-induced B cell antigen receptor signaling viewed in living cells by fluorescence resonance energy transfer. Nat Immunol. 2005; 6(11):1168-76. DOI: 10.1038/ni1262. View

Bacia K, Majoul I, Schwille P . Probing the endocytic pathway in live cells using dual-color fluorescence cross-correlation analysis. Biophys J. 2002; 83(2):1184-93. PMC: 1302220. DOI: 10.1016/S0006-3495(02)75242-9. View

Demarco I, Periasamy A, Booker C, Day R . Monitoring dynamic protein interactions with photoquenching FRET. Nat Methods. 2006; 3(7):519-24. PMC: 2921620. DOI: 10.1038/nmeth889. View

Bacia K, Kim S, Schwille P . Fluorescence cross-correlation spectroscopy in living cells. Nat Methods. 2006; 3(2):83-9. DOI: 10.1038/nmeth822. View

10.

Rocheleau J, Edidin M, Piston D . Intrasequence GFP in class I MHC molecules, a rigid probe for fluorescence anisotropy measurements of the membrane environment. Biophys J. 2003; 84(6):4078-86. PMC: 1302987. DOI: 10.1016/S0006-3495(03)75133-9. View

11.

Rizzo M, Piston D . High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy. Biophys J. 2004; 88(2):L14-6. PMC: 1305173. DOI: 10.1529/biophysj.104.055442. View

12.

Tramier M, Zahid M, Mevel J, Masse M, Coppey-Moisan M . Sensitivity of CFP/YFP and GFP/mCherry pairs to donor photobleaching on FRET determination by fluorescence lifetime imaging microscopy in living cells. Microsc Res Tech. 2006; 69(11):933-9. DOI: 10.1002/jemt.20370. View

13.

Lamb D, Muller B, Brauchle C . Enhancing the sensitivity of fluorescence correlation spectroscopy by using time-correlated single photon counting. Curr Pharm Biotechnol. 2005; 6(5):405-14. DOI: 10.2174/138920105774370625. View

14.

Kuroyama H, Ikeda T, Kasai M, Yamasaki S, Tatsumi M, Utsuyama M . Identification of a novel isoform of ZAP-70, truncated ZAP kinase. Biochem Biophys Res Commun. 2004; 315(4):935-41. DOI: 10.1016/j.bbrc.2004.01.127. View

15.

Krause C, Mei E, Mirochnitchenko O, Lavnikova N, Xie J, Jia Y . Interactions among the components of the interleukin-10 receptor complex. Biochem Biophys Res Commun. 2005; 340(2):377-85. DOI: 10.1016/j.bbrc.2005.11.182. View

16.

Wallrabe H, Stanley M, Periasamy A, Barroso M . One- and two-photon fluorescence resonance energy transfer microscopy to establish a clustered distribution of receptor-ligand complexes in endocytic membranes. J Biomed Opt. 2003; 8(3):339-46. DOI: 10.1117/1.1584444. View

17.

Zal T, Gascoigne N . Photobleaching-corrected FRET efficiency imaging of live cells. Biophys J. 2004; 86(6):3923-39. PMC: 1304294. DOI: 10.1529/biophysj.103.022087. View

18.

Vamosi G, Bodnar A, Vereb G, Jenei A, Goldman C, Langowski J . IL-2 and IL-15 receptor alpha-subunits are coexpressed in a supramolecular receptor cluster in lipid rafts of T cells. Proc Natl Acad Sci U S A. 2004; 101(30):11082-7. PMC: 503744. DOI: 10.1073/pnas.0403916101. View

19.

Kohl T, Heinze K, Kuhlemann R, Koltermann A, Schwille P . A protease assay for two-photon crosscorrelation and FRET analysis based solely on fluorescent proteins. Proc Natl Acad Sci U S A. 2002; 99(19):12161-6. PMC: 129415. DOI: 10.1073/pnas.192433499. View

20.

Clayton A, Hanley Q, Verveer P . Graphical representation and multicomponent analysis of single-frequency fluorescence lifetime imaging microscopy data. J Microsc. 2003; 213(1):1-5. DOI: 10.1111/j.1365-2818.2004.01265.x. View