A dansyl fluorescence-based assay for monitoring kinetics of lipid extraction and transfer.
Lipid transfer proteins have important roles in cellular biology, and fluorescence spectroscopy has found wide range use as a facile means for time-resolved monitoring of protein-lipid interactions. Here, we show how the fluorescence emission properties of dansyl-DHPE can be exploited to characterize lipid extraction and lipid transfer kinetics. The GM2 activator protein serves as an example of a lipid transfer protein where the ability to independently characterize lipid extraction from donor vesicles, formation of a protein:lipid complex in solution, and release of lipid from the complex to acceptor liposomes is crucial for full kinetic characterization of lipid transfer.
Ran
Yong
Y
Department of Chemistry, University of Florida, P.O. Box 117200, Gainesville, FL 32611-7200, USA.
Fanucci
Gail E
GE
D001681:Biological Assay; D004735:Energy Transfer; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D049289:G(M2) Activator Protein / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008055:Lipids / Q000302:isolation & purification*; D010713:Phosphatidylcholines / Q000737:chemistry* / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence
C496906:1-myristoyl-2-(12-((5-dimethylamino-1-naphthalenesulfonyl)amino)dodecanoyl)-sn-glycero-3-phosphocholine; D005456:Fluorescent Dyes; D049289:G(M2) Activator Protein; D008055:Lipids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; C007510:1,2-dipalmitoyl-3-phosphatidylethanolamine