Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose
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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2021-07-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/65417 |
| _version_ | 1818026631963344896 |
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| author | Cheng Qian Zhaofa Wu Rongbo Sun Huasheng Yu Jianzhi Zeng Yi Rao Yulong Li |
| author_facet | Cheng Qian Zhaofa Wu Rongbo Sun Huasheng Yu Jianzhi Zeng Yi Rao Yulong Li |
| author_sort | Cheng Qian |
| collection | DOAJ |
| description | 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 seven transporters in overlap. Ultrastructural analysis further supported that 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. |
| first_indexed | 2024-12-10T04:35:05Z |
| format | Article |
| id | doaj.art-4102e0d2badc4b2089608a6b3379cab9 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T04:35:05Z |
| publishDate | 2021-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-4102e0d2badc4b2089608a6b3379cab92022-12-22T02:02:01ZengeLife Sciences Publications LtdeLife2050-084X2021-07-011010.7554/eLife.65417Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucoseCheng Qian0https://orcid.org/0000-0002-9926-4406Zhaofa Wu1https://orcid.org/0000-0003-2027-5194Rongbo Sun2https://orcid.org/0000-0002-4364-8435Huasheng Yu3https://orcid.org/0000-0001-5641-2512Jianzhi Zeng4https://orcid.org/0000-0002-5380-6281Yi Rao5https://orcid.org/0000-0002-0405-5426Yulong Li6https://orcid.org/0000-0002-9166-9919School of Life Sciences, Tsinghua University, Beijing, ChinaState Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China; PKU-IDG/McGovern Institute for Brain Research, Beijing, ChinaState Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China; PKU-IDG/McGovern Institute for Brain Research, Beijing, ChinaState Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China; PKU-IDG/McGovern Institute for Brain Research, Beijing, China; Peking-Tsinghua Center for Life Sciences, Beijing, ChinaState Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China; PKU-IDG/McGovern Institute for Brain Research, Beijing, China; Peking-Tsinghua Center for Life Sciences, Beijing, ChinaState Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China; PKU-IDG/McGovern Institute for Brain Research, Beijing, China; Peking-Tsinghua Center for Life Sciences, Beijing, China; Chinese Institute for Brain Research, Beijing, ChinaState Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China; PKU-IDG/McGovern Institute for Brain Research, Beijing, China; Peking-Tsinghua Center for Life Sciences, Beijing, China; Chinese Institute for Brain Research, Beijing, ChinaVesicular 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 seven transporters in overlap. Ultrastructural analysis further supported that 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.https://elifesciences.org/articles/65417neurotransmitterstransportersolute carriersynaptic vesiclesUDP-glucose |
| spellingShingle | Cheng Qian Zhaofa Wu Rongbo Sun Huasheng Yu Jianzhi Zeng Yi Rao Yulong Li Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose eLife neurotransmitters transporter solute carrier synaptic vesicles UDP-glucose |
| title | Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose |
| title_full | Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose |
| title_fullStr | Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose |
| title_full_unstemmed | Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose |
| title_short | Localization, proteomics, and metabolite profiling reveal a putative vesicular transporter for UDP-glucose |
| title_sort | localization proteomics and metabolite profiling reveal a putative vesicular transporter for udp glucose |
| topic | neurotransmitters transporter solute carrier synaptic vesicles UDP-glucose |
| url | https://elifesciences.org/articles/65417 |
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