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FGF21 trafficking in intact human cells revealed by cryo-electron tomography with gold nanoparticles

Azubel, Maia and Carter, Stephen D. and Weiszmann, Jennifer and Zhang, Jun and Jensen, Grant J. and Li, Yang and Kornberg, Roger D. (2019) FGF21 trafficking in intact human cells revealed by cryo-electron tomography with gold nanoparticles. eLife, 8 . Art. No. e43146. ISSN 2050-084X. PMCID PMC6349402. http://resolver.caltech.edu/CaltechAUTHORS:20190207-143318376

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Abstract

The fibroblast growth factor FGF21 was labeled with molecularly defined gold nanoparticles (AuNPs), applied to human adipocytes, and imaged by cryo-electron tomography (cryo-ET). Most AuNPs were in pairs about 80 Å apart, on the outer cell surface. Pairs of AuNPs were also abundant inside the cells in clathrin-coated vesicles and endosomes. AuNPs were present but no longer paired in multivesicular bodies. FGF21 could thus be tracked along the endocytotic pathway. The methods developed here to visualize signaling coupled to endocytosis can be applied to a wide variety of cargo and may be extended to studies of other intracellular transactions.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.7554/elife.43146DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6349402PubMed CentralArticle
https://doi.org/10.7554/eLife.43146.015DOITransparent reporting form
ORCID:
AuthorORCID
Azubel, Maia0000-0002-1584-2695
Carter, Stephen D.0000-0002-4237-4276
Zhang, Jun0000-0003-4154-3741
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2019 Azubel et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Received: 26 October 2018; Accepted: 07 January 2019; Published: 28 January 2019. Data availability: All data is provided in the manuscript and supporting files. This research was supported by NIH grants AI 21144 to RDK and R35 122588 to GJJ. We thank Amgen Department of Protein Sciences for providing some of the reagents used in this study. We thank Dr. E P Geiduschek for discussion and comments on the manuscript. The funders had role in study design, and the decision to submit the work for publication. Competing interests: Jennifer Weiszmann, Jun Zhang, Yang Li: Employee of Amgen at the time the study was conducted. There are no other competing financial interests to declare. The other authors declare that no competing interests exist. Author contributions: Maia Azubel, Conceptualization, Data curation, Formal analysis, Supervision, Validation, Investigation, Visualization, Methodology, Writing—original draft, Project administration, Writing—review and editing; Stephen D Carter, Investigation, Methodology, Writing—review and editing; Jennifer Weiszmann, Resources, Investigation, Visualization, Methodology, Writing—review and editing; Jun Zhang, Resources, Writing—review and editing; Grant J Jensen, Conceptualization, Resources, Funding acquisition, Writing—review and editing; Yang Li, Conceptualization, Resources, Supervision, Funding acquisition, Methodology, Writing—review and editing; Roger D Kornberg, Conceptualization, Resources, Supervision, Writing—original draft, Writing—review and editing.
Funders:
Funding AgencyGrant Number
NIHAI 21144
NIHR35 122588
PubMed Central ID:PMC6349402
Record Number:CaltechAUTHORS:20190207-143318376
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20190207-143318376
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:92767
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:08 Feb 2019 00:19
Last Modified:08 Feb 2019 00:19

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