1. Neuroscience
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Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose

  1. Cheng Qian
  2. Zhaofa Wu
  3. Rongbo Sun
  4. Huasheng Yu
  5. Jianzhi Zeng
  6. Yi Rao
  7. Yulong Li  Is a corresponding author
  1. School of Life Sciences, Tsinghua University, China
  2. Peking University School of Life Sciences, China
  3. Peking University, China
  4. Peiking University, China
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Cite this article as: eLife 2021;10:e65417 doi: 10.7554/eLife.65417

Abstract

Vesicular neurotransmitter transporters (VNTs) mediate the selective uptake and enrichment of small molecule neurotransmitters into synaptic vesicles (SVs) and are therefore a major determinant of the synaptic output of specific neurons. To identify novel VNTs expressed on SVs (thus identifying new neurotransmitters and/or neuromodulators), we conducted localization profiling of 361 solute carrier (SLC) transporters tagging with a fluorescent protein in neurons, which revealed 40 possible candidates through comparison with a known SV marker. We parallelly performed proteomics analysis of immunoisolated SVs and identified 7 transporters in overlap. Ultrastructural analysis confirmed one of the transporters, SLC35D3, localized to SVs. Finally, by combining metabolite profiling with a radiolabeled substrate transport assay, we identified UDP-glucose as the principal substrate for SLC35D3. These results provide new insights into the functional role of SLC transporters in neurotransmission and improve our understanding of the molecular diversity of chemical transmitters.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Cheng Qian

    School of Life Sciences, School of Life Sciences, Tsinghua University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Zhaofa Wu

    State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Rongbo Sun

    State Key Laboratory of Membrane Biology, School of Life Sciences, Peking University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Huasheng Yu

    Peking-Tsinghua Center for Life Sciences, Peiking University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Jianzhi Zeng

    Peking-Tsinghua Center for Life Sciences, Peiking University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Yi Rao

    Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0405-5426
  7. Yulong Li

    State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
    For correspondence
    yulongli@pku.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9166-9919

Funding

Beijing Municipal Science & Technology Commission (Z181100001318002)

  • Yulong Li

Peking-Tsinghua Center for Life Sciences

  • Yulong Li

State Key Laboratory of Membrane Biology

  • Yulong Li

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal procedures were performed using protocols approved by the Institutional Animal Care and Use Committee at Peking University. ( LSC LiYL 1 )

Reviewing Editor

  1. Rebecca Seal, University of Pittsburgh School of Medicine, United States

Publication history

  1. Received: December 3, 2020
  2. Accepted: July 15, 2021
  3. Accepted Manuscript published: July 16, 2021 (version 1)
  4. Version of Record published: August 18, 2021 (version 2)

Copyright

© 2021, Qian et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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