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Nanogold as a specific marker for electron cryotomography

He, Yongning and Jensen, Grant J. and Bjorkman, Pamela J. (2009) Nanogold as a specific marker for electron cryotomography. Microscopy and Microanalysis, 15 (3). pp. 183-188. ISSN 1431-9276. PMCID PMC2785728. https://resolver.caltech.edu/CaltechAUTHORS:20090813-075852803

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1017/S1431927609090424DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2785728PubMed CentralArticle
ORCID:
AuthorORCID
Jensen, Grant J.0000-0003-1556-4864
Bjorkman, Pamela J.0000-0002-2277-3990
Additional Information:© 2009 Microscopy Society of America. Received December 17, 2008; accepted April 3, 2009. We thank Noreen Tiangco for preparing Nanogold-Fc, William Tivol for the help with microscopy, and members of the Bjorkman and Jensen laboratories for helpful suggestions. This work was supported by a postdoctoral fellowship from the Cancer Research Institute (Y.H.), the National Institutes of Health (2 R37 AI041239-06A1 to P.J.B.), and gifts to Caltech to support electron microscopy from the Gordon and Betty Moore Foundation and the Agouron Institute.
Funders:
Funding AgencyGrant Number
Cancer Research InstituteUNSPECIFIED
NIH2 R37 AI041239-06A1
Gordon and Betty Moore FoundationUNSPECIFIED
Agouron InstituteUNSPECIFIED
Subject Keywords:Nanogold; electron cryotomography; labeling; liposome; FcRn
Issue or Number:3
PubMed Central ID:PMC2785728
Record Number:CaltechAUTHORS:20090813-075852803
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20090813-075852803
Official Citation:He, Y., Jensen, G., & Bjorkman, P. (2009). Nanogold as a Specific Marker for Electron Cryotomography. Microscopy and Microanalysis, 15(3), 183-188. doi:10.1017/S1431927609090424
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:15003
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:13 Aug 2009 20:45
Last Modified:27 Apr 2020 17:17

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