Home LiteratureArticle Details
PMID: 16800872 Published · epublish English Journal Article

The visible touch: in planta visualization of protein-protein interactions by fluorophore-based methods.

Plant methods ·Vol. 2 ·2006-06-26 ·Pages 12

Bhat RA, Lahaye T, Panstruga R

Abstract

Non-invasive fluorophore-based protein interaction assays like fluorescence resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC, also referred to as "split YFP") have been proven invaluable tools to study protein-protein interactions in living cells. Both methods are now frequently used in the plant sciences and are likely to develop into standard techniques for the identification, verification and in-depth analysis of polypeptide interactions. In this review, we address the individual strengths and weaknesses of both approaches and provide an outlook about new directions and possible future developments for both techniques.

Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bhat Riyaz A
Department of Plant-Microbe Interactions, Max-Planck-Institute for Plant Breeding Research, Carl-von-Linné-Weg 10, D-50829 Köln, Germany. bhat@mpiz-koeln.mpg.de
Lahaye Thomas
Panstruga Ralph
References (88)
88 references, click to expand
  1. Imaging protein-protein interactions in living cells.
    Plant Mol Biol. 2002 Dec;50(6):871-83 PMID: 12516859
  2. Nuclear import of the transcription factor SHOOT MERISTEMLESS depends on heterodimerization with BLH proteins expressed in discrete sub-domains of the shoot apical meristem of Arabidopsis thaliana.
    Nucleic Acids Res. 2006;34(4):1281-92 PMID: 16513846
  3. Simultaneous visualization of multiple protein interactions in living cells using multicolor fluorescence complementation analysis.
    Nat Biotechnol. 2003 May;21(5):539-45 PMID: 12692560
  4. Analysis of MADS box protein-protein interactions in living plant cells.
    Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2416-21 PMID: 11854533
  5. Fluorescence resonance energy transfer (FRET) microscopy imaging of live cell protein localizations.
    J Cell Biol. 2003 Mar 3;160(5):629-33 PMID: 12615908
  6. Fluorescence resonance energy transfer.
    Curr Opin Biotechnol. 1995 Feb;6(1):103-10 PMID: 7534502
  7. A cDNA library functional screening strategy based on fluorescent protein complementation assays to identify novel components of signaling pathways.
    Methods. 2004 Apr;32(4):381-8 PMID: 15003600
  8. The plant VirE2 interacting protein 1. a molecular link between the Agrobacterium T-complex and the host cell chromatin?
    Plant Physiol. 2005 Jul;138(3):1318-21 PMID: 16010006
  9. LIP19, a basic region leucine zipper protein, is a Fos-like molecular switch in the cold signaling of rice plants.
    Plant Cell Physiol. 2005 Oct;46(10):1623-34 PMID: 16051676
  10. Blue-native PAGE in plants: a tool in analysis of protein-protein interactions.
    Plant Methods. 2005;1(1):11 PMID: 16287510
  11. Visualization of the spatial and temporal dynamics of intracellular signaling.
    Dev Cell. 2003 Mar;4(3):295-305 PMID: 12636912
  12. Reassembled GFP: detecting protein-protein interactions and protein expression patterns.
    Methods Biochem Anal. 2006;47:391-405 PMID: 16342418
  13. FD, a bZIP protein mediating signals from the floral pathway integrator FT at the shoot apex.
    Science. 2005 Aug 12;309(5737):1052-6 PMID: 16099979
  14. Arabidopsis SENESCENCE-ASSOCIATED GENE101 stabilizes and signals within an ENHANCED DISEASE SUSCEPTIBILITY1 complex in plant innate immunity.
    Plant Cell. 2005 Sep;17(9):2601-13 PMID: 16040633
  15. In planta analysis of protein-protein interactions related to light signaling by bimolecular fluorescence complementation.
    Protoplasma. 2005 Dec;226(3-4):137-46 PMID: 16333572
  16. Three-chromophore FRET microscopy to analyze multiprotein interactions in living cells.
    Nat Methods. 2004 Dec;1(3):209-17 PMID: 15782196
  17. The renaissance of fluorescence resonance energy transfer.
    Nat Struct Biol. 2000 Sep;7(9):730-4 PMID: 10966639
  18. Green fluorescent protein as a scaffold for intracellular presentation of peptides.
    Nucleic Acids Res. 1998 Jan 15;26(2):623-30 PMID: 9421525
  19. The VirE3 protein of Agrobacterium mimics a host cell function required for plant genetic transformation.
    EMBO J. 2005 Jan 26;24(2):428-37 PMID: 15616576
  20. Green fluorescent protein as a marker for gene expression.
    Science. 1994 Feb 11;263(5148):802-5 PMID: 8303295
  21. Probing plasma membrane microdomains in cowpea protoplasts using lipidated GFP-fusion proteins and multimode FRET microscopy.
    J Microsc. 2004 May;214(Pt 2):190-200 PMID: 15102066
  22. Bimolecular fluorescence complementation analysis of cytochrome p450 2c2, 2e1, and NADPH-cytochrome p450 reductase molecular interactions in living cells.
    Drug Metab Dispos. 2005 Sep;33(9):1382-90 PMID: 15980100
  23. A draft sequence of the rice genome (Oryza sativa L. ssp. japonica).
    Science. 2002 Apr 5;296(5565):92-100 PMID: 11935018
  24. Fluorescence lifetime imaging microscopy (FLIM).
    Adv Biochem Eng Biotechnol. 2005;95:143-75 PMID: 16080268
  25. Design of the linkers which effectively separate domains of a bifunctional fusion protein.
    Protein Eng. 2001 Aug;14(8):529-32 PMID: 11579220
  26. Single-molecule spectroscopic methods.
    Curr Opin Struct Biol. 2004 Oct;14(5):531-40 PMID: 15465312
  27. A guide to choosing fluorescent proteins.
    Nat Methods. 2005 Dec;2(12):905-9 PMID: 16299475
  28. Fluorescence resonance energy transfer from cyan to yellow fluorescent protein detected by acceptor photobleaching using confocal microscopy and a single laser.
    J Microsc. 2003 Jan;209(Pt 1):56-70 PMID: 12535185
  29. Studying protein-protein interactions via blot overlay or Far Western blot.
    Methods Mol Biol. 2004;261:167-74 PMID: 15064457
  30. Efficiently folding and circularly permuted variants of the Sapphire mutant of GFP.
    BMC Biotechnol. 2003 May 22;3:5 PMID: 12769828
  31. Recruitment and interaction dynamics of plant penetration resistance components in a plasma membrane microdomain.
    Proc Natl Acad Sci U S A. 2005 Feb 22;102(8):3135-40 PMID: 15703292
  32. The green fluorescent protein.
    Annu Rev Biochem. 1998;67:509-44 PMID: 9759496
  33. Chloroplast division site placement requires dimerization of the ARC11/AtMinD1 protein in Arabidopsis.
    J Cell Sci. 2004 May 1;117(Pt 11):2399-410 PMID: 15126639
  34. A draft sequence of the rice genome (Oryza sativa L. ssp. indica).
    Science. 2002 Apr 5;296(5565):79-92 PMID: 11935017
  35. Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation.
    Plant J. 2004 Nov;40(3):428-38 PMID: 15469500
  36. Plant immunity: the origami of receptor activation.
    Curr Biol. 2004 Jan 6;14(1):R22-4 PMID: 14711430
  37. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein.
    Nat Biotechnol. 2004 Dec;22(12):1567-72 PMID: 15558047
  38. FRET imaging.
    Nat Biotechnol. 2003 Nov;21(11):1387-95 PMID: 14595367
  39. Primary structure of the Aequorea victoria green-fluorescent protein.
    Gene. 1992 Feb 15;111(2):229-33 PMID: 1347277
  40. A central role of Arabidopsis thaliana ovate family proteins in networking and subcellular localization of 3-aa loop extension homeodomain proteins.
    Proc Natl Acad Sci U S A. 2005 Mar 29;102(13):4908-12 PMID: 15781858
  41. Measuring fast dynamics in solutions and cells with a laser scanning microscope.
    Biophys J. 2005 Aug;89(2):1317-27 PMID: 15908582
  42. Calmodulin interacts with MLO protein to regulate defence against mildew in barley.
    Nature. 2002 Mar 28;416(6879):447-51 PMID: 11919636
  43. In situ analysis of spatial relationships between proteins of the nuclear pore complex.
    Biophys J. 2002 Dec;83(6):3626-36 PMID: 12496130
  44. Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements.
    FEBS Lett. 2005 Aug 15;579(20):4374-82 PMID: 16061227
  45. Spectral profiling for the simultaneous observation of four distinct fluorescent proteins and detection of protein-protein interaction via fluorescence resonance energy transfer in tobacco leaf nuclei.
    Plant Physiol. 2002 Jul;129(3):931-42 PMID: 12114548
  46. GFP-based FRET microscopy in living plant cells.
    Trends Plant Sci. 1999 Jul;4(7):287-291 PMID: 10407445
  47. Nanoscale organization of multiple GPI-anchored proteins in living cell membranes.
    Cell. 2004 Feb 20;116(4):577-89 PMID: 14980224
  48. FRET or no FRET: a quantitative comparison.
    Biophys J. 2003 Jun;84(6):3992-4010 PMID: 12770904
  49. Fluorescence-quenching and resonance energy transfer studies of lipid microdomains in model and biological membranes.
    Mol Membr Biol. 2006 Jan-Feb;23(1):5-16 PMID: 16611577
  50. Photoconversion of YFP into a CFP-like species during acceptor photobleaching FRET experiments.
    Nat Methods. 2005 Nov;2(11):801 PMID: 16278647
  51. Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells.
    Science. 2002 May 3;296(5569):913-6 PMID: 11988576
  52. Assessment of the integral membrane protein topology in living cells.
    Plant J. 2006 Apr;46(1):145-54 PMID: 16553902
  53. Interaction of maize Opaque-2 and the transcriptional co-activators GCN5 and ADA2, in the modulation of transcriptional activity.
    Plant Mol Biol. 2004 May;55(2):239-52 PMID: 15604678
  54. Determination of tumor necrosis factor receptor-associated factor trimerization in living cells by CFP->YFP->mRFP FRET detected by flow cytometry.
    Nucleic Acids Res. 2005;33(6):e61 PMID: 15805120
  55. Evolutionary optimization of fluorescent proteins for intracellular FRET.
    Nat Biotechnol. 2005 Mar;23(3):355-60 PMID: 15696158
  56. Fluorescence cross-correlation spectroscopy in living cells.
    Nat Methods. 2006 Feb;3(2):83-9 PMID: 16432516
  57. Conserved extracellular cysteine residues and cytoplasmic loop-loop interplay are required for functionality of the heptahelical MLO protein.
    Biochem J. 2005 Jan 1;385(Pt 1):243-54 PMID: 15352871
  58. Reliable and global measurement of fluorescence resonance energy transfer using fluorescence microscopes.
    Biophys J. 2001 Oct;81(4):2395-402 PMID: 11566809
  59. Plastid division is mediated by combinatorial assembly of plastid division proteins.
    Plant J. 2005 Sep;43(6):811-23 PMID: 16146521
  60. Visualization of maltose uptake in living yeast cells by fluorescent nanosensors.
    Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9846-51 PMID: 12097642
  61. Improvement of a FRET-based indicator for cAMP by linker design and stabilization of donor-acceptor interaction.
    J Mol Biol. 2005 Dec 2;354(3):546-55 PMID: 16257413
  62. Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions.
    Biotechniques. 2006 Jan;40(1):61-6 PMID: 16454041
  63. The Arabidopsis kinase-associated protein phosphatase controls internalization of the somatic embryogenesis receptor kinase 1.
    Genes Dev. 2002 Jul 1;16(13):1707-20 PMID: 12101128
  64. Label-free screening of bio-molecular interactions.
    Anal Bioanal Chem. 2003 Nov;377(5):834-42 PMID: 12904946
  65. Uncoupling of the functions of the Arabidopsis VIP1 protein in transient and stable plant genetic transformation by Agrobacterium.
    Proc Natl Acad Sci U S A. 2005 Apr 19;102(16):5733-8 PMID: 15824315
  66. Creating new fluorescent probes for cell biology.
    Nat Rev Mol Cell Biol. 2002 Dec;3(12):906-18 PMID: 12461557
  67. The organization of the core proteins of the yeast spindle pole body.
    Mol Biol Cell. 2005 Jul;16(7):3341-52 PMID: 15872084
  68. Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin.
    Nature. 1997 Aug 28;388(6645):882-7 PMID: 9278050
  69. Co-immunoprecipitation from transfected cells.
    Methods Mol Biol. 2004;261:337-50 PMID: 15064468
  70. The DOF protein, SAD, interacts with GAMYB in plant nuclei and activates transcription of endosperm-specific genes during barley seed development.
    Plant J. 2005 Jun;42(5):652-62 PMID: 15918880
  71. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana.
    Nature. 2000 Dec 14;408(6814):796-815 PMID: 11130711
  72. Analysing protein-protein interactions with the yeast two-hybrid system.
    Plant Mol Biol. 2002 Dec;50(6):855-70 PMID: 12516858
  73. Illuminating insights into protein-protein interactions using bioluminescence resonance energy transfer (BRET).
    Nat Methods. 2006 Mar;3(3):165-74 PMID: 16489332
  74. Subcellular localization and oligomerization of the Arabidopsis thaliana somatic embryogenesis receptor kinase 1 protein.
    J Mol Biol. 2001 Jun 8;309(3):641-55 PMID: 11397085
  75. An improved cyan fluorescent protein variant useful for FRET.
    Nat Biotechnol. 2004 Apr;22(4):445-9 PMID: 14990965
  76. Energy transfer: a spectroscopic ruler.
    Proc Natl Acad Sci U S A. 1967 Aug;58(2):719-26 PMID: 5233469
  77. In vivo imaging of MADS-box transcription factor interactions.
    J Exp Bot. 2006;57(1):33-42 PMID: 16291798
  78. New tools for in vivo fluorescence tagging.
    Curr Opin Plant Biol. 2005 Dec;8(6):565-73 PMID: 16188488
  79. Using GFP in FRET-based applications.
    Trends Cell Biol. 1999 Feb;9(2):57-60 PMID: 10087619
  80. Quantitative fluorescence resonance energy transfer measurements using fluorescence microscopy.
    Biophys J. 1998 May;74(5):2702-13 PMID: 9591694
  81. Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation.
    Mol Cell. 2002 Apr;9(4):789-98 PMID: 11983170
  82. Using intrinsically fluorescent proteins for plant cell imaging.
    Plant J. 2006 Feb;45(4):599-615 PMID: 16441351
  83. Detection of protein-protein interactions in plants using bimolecular fluorescence complementation.
    Plant J. 2004 Nov;40(3):419-27 PMID: 15469499
  84. Circular permutation and receptor insertion within green fluorescent proteins.
    Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11241-6 PMID: 10500161
  85. G Protein activation without subunit dissociation depends on a G{alpha}(i)-specific region.
    J Biol Chem. 2005 Jul 1;280(26):24584-90 PMID: 15866880
  86. Involvement of targeted proteolysis in plant genetic transformation by Agrobacterium.
    Nature. 2004 Sep 2;431(7004):87-92 PMID: 15343337
  87. Functional interaction of phytochrome B and cryptochrome 2.
    Nature. 2000 Nov 9;408(6809):207-11 PMID: 11089975
  88. Detecting protein-protein interactions with a green fluorescent protein fragment reassembly trap: scope and mechanism.
    J Am Chem Soc. 2005 Jan 12;127(1):146-57 PMID: 15631464
Article Info
Journal
Plant methods
Abbr.
Plant Methods
ISSN
1746-4811
DOI
10.1186/1746-4811-2-12
Email
bhat@mpiz-koeln.mpg.de
Published
2006-06-26
Epub
2006-00-26
Pages
12
Language
English
Region
England
NLM ID
101245798
PMCID
PMC1523328
External Links
PubMed source DOI: 10.1186/1746-4811-2-12 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2