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Nanogold as a Specific Marker for Electron Cryotomography

Published online by Cambridge University Press:  22 May 2009

Yongning He ,
Grant J. Jensen  and
Pamela J. Bjorkman
Yongning He
Affiliation:
Division of Biology, California Institute of Technology, 114-96, 1200 East California Blvd., Pasadena, CA 91125
Grant J. Jensen
Affiliation:
Division of Biology, California Institute of Technology, 114-96, 1200 East California Blvd., Pasadena, CA 91125 Howard Hughes Medical Institute, California Institute of Technology, 1200 East California Blvd., Pasadena, CA 91125
Pamela J. Bjorkman*
Affiliation:
Division of Biology, California Institute of Technology, 114-96, 1200 East California Blvd., Pasadena, CA 91125 Howard Hughes Medical Institute, California Institute of Technology, 1200 East California Blvd., Pasadena, CA 91125
*
Corresponding author. E-mail: bjorkman@caltech.edu
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Abstract

While electron cryotomography (ECT) provides “molecular” resolution, three-dimensional images of unique biological specimens, sample crowdedness, and/or resolution limitations can make it difficult to identify specific macromolecular components. Here we used a 1.4 nm Nanogold® cluster specifically attached to the Fc fragment of IgG to monitor its interaction with the neonatal Fc receptor (FcRn), a membrane-bound receptor that transports IgG across cells in acidic intracellular vesicles. ECT was used to image complexes formed by Nanogold-labeled Fc bound to FcRn attached to the outer surface of synthetic liposomes. In the resulting three-dimensional reconstructions, 1.4 nm Nanogold particles were distributed predominantly along the interfaces where 2:1 FcRn-Fc complexes bridged adjacent lipid bilayers. These results demonstrate that the 1.4 nm Nanogold cluster is visible in tomograms of typically thick samples (∼250 nm) recorded with defocuses appropriate for large macromolecules and is thus an effective marker.

Keywords

Nanogoldelectron cryotomographylabelingliposomeFcRn
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 15 , Issue 3 , June 2009 , pp. 183 - 188
Copyright
Copyright © Microscopy Society of America 2009

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