Biomaterial Database

Monitoring Transmembrane and Peripheral Membrane Protein Interactions by Förster Resonance Energy Transfer Using Fluorescence Lifetime Imaging Microscopy. 2022

Janhavi Nagwekar, and Caterina Di Ciano-Oliveira, and Gregory D Fairn

Keenan Research Centre for Biomedical Science, St. Michael's Hospital, Unity Health Toronto, Toronto, ON, Canada.

Caveolae are bulb-shaped invaginations of the plasma membrane that are enriched in specific lipids including cholesterol, phosphatidylserine and sphingolipids. Caveolae have many described cellular roles and functions, including endocytic transport, transcytosis, mechanosensing, and serving as a buffer against plasmalemmal stress. Caveola are formed through interactions between integral membrane proteins (Caveolin) and a cavin family of peripheral proteins (Cavins). Nearly half of the human proteome resides within or at the surface of membranes. Studying protein-protein interactions, especially of transmembrane domain containing proteins can be challenging. Fortunately, sophisticated biophysical methods allow for the monitoring of protein interactions in intact cells. Here, we describe the principles of Förster resonance energy transfer, fluorescence lifetime, and how their properties can be used to assess protein-protein interactions. Additionally, we discuss and demonstrate how fluorescence lifetime can be monitored microscopically thereby providing caveolin-cavin interaction data from living cells.

UI	MeSH Term	Description	Entries
D008565	Membrane Proteins	Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.	Cell Membrane Protein,Cell Membrane Proteins,Cell Surface Protein,Cell Surface Proteins,Integral Membrane Proteins,Membrane-Associated Protein,Surface Protein,Surface Proteins,Integral Membrane Protein,Membrane Protein,Membrane-Associated Proteins,Membrane Associated Protein,Membrane Associated Proteins,Membrane Protein, Cell,Membrane Protein, Integral,Membrane Proteins, Integral,Protein, Cell Membrane,Protein, Cell Surface,Protein, Integral Membrane,Protein, Membrane,Protein, Membrane-Associated,Protein, Surface,Proteins, Cell Membrane,Proteins, Cell Surface,Proteins, Integral Membrane,Proteins, Membrane,Proteins, Membrane-Associated,Proteins, Surface,Surface Protein, Cell
D008853	Microscopy	The use of instrumentation and techniques for visualizing material and details that cannot be seen by the unaided eye. It is usually done by enlarging images, transmitted by light or electron beams, with optical or magnetic lenses that magnify the entire image field. With scanning microscopy, images are generated by collecting output from the specimen in a point-by-point fashion, on a magnified scale, as it is scanned by a narrow beam of light or electrons, a laser, a conductive probe, or a topographical probe.	Compound Microscopy,Hand-Held Microscopy,Light Microscopy,Optical Microscopy,Simple Microscopy,Hand Held Microscopy,Microscopy, Compound,Microscopy, Hand-Held,Microscopy, Light,Microscopy, Optical,Microscopy, Simple
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D016601	RNA-Binding Proteins	Proteins that bind to RNA molecules. Included here are RIBONUCLEOPROTEINS and other proteins whose function is to bind specifically to RNA.	Double-Stranded RNA-Binding Protein,Double-Stranded RNA-Binding Proteins,ds RNA-Binding Protein,RNA-Binding Protein,ds RNA-Binding Proteins,Double Stranded RNA Binding Protein,Double Stranded RNA Binding Proteins,Protein, Double-Stranded RNA-Binding,Protein, ds RNA-Binding,RNA Binding Protein,RNA Binding Proteins,RNA-Binding Protein, Double-Stranded,RNA-Binding Protein, ds,RNA-Binding Proteins, Double-Stranded,ds RNA Binding Protein
D051242	Caveolin 1	A tyrosine phosphoprotein that plays an essential role in CAVEOLAE formation. It binds CHOLESTEROL and is involved in LIPIDS transport, membrane traffic, and SIGNAL TRANSDUCTION.	Caveolin-1,VIP21 Protein,Vesicular Integral Membrane Protein 21 kDa,alpha-Caveolin,beta-Caveolin,alpha Caveolin,beta Caveolin
D031541	Fluorescence Resonance Energy Transfer	A type of FLUORESCENCE SPECTROSCOPY using two FLUORESCENT DYES with overlapping emission and absorption spectra, which is used to indicate proximity of labeled molecules. This technique is useful for studying interactions of molecules and PROTEIN FOLDING.	Forster Resonance Energy Transfer