Combined Bimolecular Fluorescence Complementation and Forster Resonance Energy Transfer Reveals Ternary SNARE Complex Formation in Living Plant Cells

被引：46

作者：

Kwaaitaal, Mark ^{[1
]}

Keinath, Nana F. ^{[1
]}

Pajonk, Simone ^{[1
]}

Biskup, Christoph ^{[2
]}

Panstruga, Ralph ^{[1
]}

机构：

[1] Max Planck Inst Plant Breeding Res, D-50829 Cologne, Germany

[2] Univ Klinikum Jena, D-07740 Jena, Germany

来源：

PLANT PHYSIOLOGY | 2010年 / 152卷 / 03期

关键词：

PROTEIN-PROTEIN INTERACTIONS; PENETRATION RESISTANCE; DISEASE-RESISTANCE; PEN1; SYNTAXIN; FRET; VISUALIZATION; BIFC; LIFETIME; CYAN; MICROSCOPY;

D O I：

10.1104/pp.109.151142

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

Various fluorophore-based microscopic methods, comprising Forster resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC), are suitable to study pairwise interactions of proteins in living cells. The analysis of interactions between more than two protein partners using these methods, however, remains difficult. In this study, we report the successful application of combined BiFC-FRET-fluorescence lifetime imaging microscopy and BiFC-FRET-acceptor photobleaching measurements to visualize the formation of ternary soluble N-ethylmaleimide-sensitive factor attachment receptor complexes in leaf epidermal cells. This method expands the repertoire of techniques to study protein-protein interactions in living plant cells by a procedure capable of visualizing simultaneously interactions between three fluorophore-tagged polypeptide partners.

引用

页码：1135 / 1147

页数：13

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