Optical single transporter recording: transport kinetics in microarrays of membrane patches.
Optical single transporter recording (OSTR) is an emerging technique for the fluorescence microscopic measurement of transport kinetics in membrane patches. Membranes are attached to transparent microarrays of cylindrical test compartments (TCs) approximately 0.1-100 mum in diameter and approximately 10-100 mum in depth. Transport across membrane patches that may contain single transporters or transporter populations is recorded by confocal microscopy. By these means transport of proteins through single nuclear pore complexes has been recorded at rates of <1 translocation/s. In addition to the high sensitivity in terms of measurable transport rates OSTR features unprecedented spatial selectivity and parallel processing. This article reviews the conceptual basis of OSTR and its realization. Applications to nuclear transport are summarized. The further development of OSTR is discussed and its extension to a diversity of transporters, including translocases and ATP-binding cassette (ABC) pumps, projected.
Peters
Reiner
R
Institut fur Medizinische Physik und Biophysik, Universitat Munster, Munster, Germany. petersr@uni-muenster.de
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D004058:Diffusion; D004867:Equipment Design; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D009038:Motion; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D009685:Nuclear Envelope / Q000378:metabolism*; D022022:Nuclear Pore / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D055095:Optics and Photonics / Q000295:instrumentation; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods; D038761:Photobleaching; D010782:Photolysis
D009687:Nuclear Proteins