title: Detection of asymmetric distribution of phospholipids by fluorescence resonance energy transfer.
abstract: It is well established that the plasma membrane exhibits an asymmetric distribution of lipids between the inner and outer leaflets of the lipid bilayer. Recent studies suggest that the asymmetric distribution changes locally and temporarily, accompanied by cellular events. However, available methods to detect lipid asymmetry lack spatio-temporal resolution. As a technique of potential use for real-time imaging of lipid asymmetry, we a novel method that utilizes fluorescence resonance energy transfer (FRET) between NBD-labeled phospholipids (donor) and extracellular rhodamine (acceptor). When cell apoptosis was induced by staurosporine, the fluorescence intensity of NBD-labeled phosphatidylserine decreased owing to FRET from NBD to rhodamine. This method provides a simple way to detect lipid asymmetry and may be useful for observing dynamic changes in asymmetric distribution of lipids.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1, Tanabe-dori, Nagoya 467-8603, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Nishimura
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tadokoro
authors.forename: Masahiko
authors.initials: M
authors.lastname: Tanaka
authors.forename: Naohide
authors.initials: N
authors.lastname: Hirashima
mesh_terms: D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000737:chemistry; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016466:CHO Cells; D006224:Cricetinae; D066246:ErbB Receptors / Q000032:analysis; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D010713:Phosphatidylcholines / Q000737:chemistry; D010718:Phosphatidylserines / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology
chemical_list: C078632:1-acyl-2-(12-((7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)dodecanoyl)phosphatidylcholine; C034296:N-(7-nitro-2,1,3-benzoxadiazol-4-yl)phosphatidylserine; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D010743:Phospholipids; D066246:ErbB Receptors; D009327:4-Chloro-7-nitrobenzofurazan; D019311:Staurosporine