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

eLife

Cheng Qian¹  Zhaofa Wu²  Rongbo Sun²  Jianzhi Zeng³  Huasheng Yu³  Yulong Li⁴  Yi Rao⁴ 

[1] School of Life Sciences, Tsinghua University, Beijing, China;State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China;PKU-IDG/McGovern Institute for Brain Research, Beijing, China;State 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;State 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, China;

关键词: neurotransmitters;    transporter;    solute carrier;    synaptic vesicles;    UDP-glucose;    Human;    Mouse;   

DOI  :  10.7554/eLife.65417

来源: eLife Sciences Publications, Ltd

【 摘 要 】

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.

【 授权许可】

CC BY