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

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Cheng Qian ◽  
Zhaofa Wu ◽  
Rongbo Sun ◽  
Huasheng Yu ◽  
Jianzhi Zeng ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Metabolite Profiling ◽  
Functional Role ◽  
Molecular Diversity ◽  
Fluorescent Protein ◽  
Proteomics Analysis ◽  
Slc Transporters ◽  
Transport Assay ◽  
Solute Carrier

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.

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

2020 ◽  
Author(s):  
Cheng Qian ◽  
Zhaofa Wu ◽  
Rongbo Sun ◽  
Huasheng Yu ◽  
Jianzhi Zeng ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Metabolite Profiling ◽  
Functional Role ◽  
Molecular Diversity ◽  
Fluorescent Protein ◽  
Proteomics Analysis ◽  
Slc Transporters ◽  
Transport Assay ◽  
Solute Carrier

AbstractVesicular 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.

scholarly journals A mammalian fatty acid hydroxylase responsible for the formation of α-hydroxylated galactosylceramide in myelin

2005 ◽  
Vol 388 (1) ◽  
pp. 245-254 ◽  
Author(s):  
Matthias ECKHARDT ◽  
Afshin YAGHOOTFAM ◽  
Simon N. FEWOU ◽  
Inge ZÖLLER ◽  
Volkmar GIESELMANN
Keyword(s):  
Fatty Acid ◽  
Functional Role ◽  
Time Course ◽  
Fluorescent Protein ◽  
Sequence Similarity ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Mammalian Brain ◽  
Fatty Acid Hydroxylase

Hydroxylation is an abundant modification of the ceramides in brain, skin, intestinal tract and kidney. Hydroxylation occurs at the sphingosine base at C-4 or within the amide-linked fatty acid. In myelin, hydroxylation of ceramide is exclusively found at the α-C atom of the fatty acid moiety. α-Hydroxylated cerebrosides are the most abundant lipids in the myelin sheath. The functional role of this modification, however, is not known. On the basis of sequence similarity to a yeast C26 fatty acid hydroxylase, we have identified a murine cDNA encoding FA2H (fatty acid 2-hydroxylase). Transfection of FA2H cDNA in CHO cells (Chinese-hamster ovary cells) led to the formation of α-hydroxylated fatty acid containing hexosylceramide. An EGFP (enhanced green fluorescent protein)–FA2H fusion protein co-localized with calnexin, indicating that the enzyme resides in the endoplasmic reticulum. FA2H is expressed in brain, stomach, skin, kidney and testis, i.e. in tissues known to synthesize fatty acid α-hydroxylated sphingolipids. The time course of its expression in brain closely follows the expression of myelin-specific genes, reaching a maximum at 2–3 weeks of age. This is in agreement with the reported time course of fatty acid α-hydroxylase activity in the developing brain. In situ hybridization of brain sections showed expression of FA2H in the white matter. Our results thus strongly suggest that FA2H is the enzyme responsible for the formation of α-hydroxylated ceramide in oligodendrocytes of the mammalian brain. Its further characterization will provide insight into the functional role of α-hydroxylation modification in myelin, skin and other organs.

scholarly journals Unraveling the functional role of the orphan solute carrier, SLC22A24 in the transport of steroid conjugates through metabolomic and genome-wide association studies

PLoS Genetics ◽  
2019 ◽  
Vol 15 (9) ◽  
pp. e1008208 ◽  
Author(s):  
Sook Wah Yee ◽  
Adrian Stecula ◽  
Huan-Chieh Chien ◽  
Ling Zou ◽  
Elena V. Feofanova ◽  
...  
Keyword(s):  
Functional Role ◽  
Association Studies ◽  
Genome Wide Association ◽  
Genome Wide Association Studies ◽  
Genome Wide ◽  
Solute Carrier ◽  
Steroid Conjugates

The role of solute carrier (SLC) transporters in actinomycin D pharmacokinetics in paediatric cancer patients

2018 ◽  
Vol 74 (12) ◽  
pp. 1575-1584 ◽  
Author(s):  
Hannah Yejin Kim ◽  
Gareth J Veal ◽  
Fanfan Zhou ◽  
Alan V Boddy
Keyword(s):  
Cancer Patients ◽  
Actinomycin D ◽  
Paediatric Cancer ◽  
Slc Transporters ◽  
Solute Carrier

scholarly journals Comparison of Cell Wall Localization among Pir Family Proteins and Functional Dissection of the Region Required for Cell Wall Binding and Bud Scar Recruitment of Pir1p

2005 ◽  
Vol 4 (11) ◽  
pp. 1872-1881 ◽  
Author(s):  
Toru Sumita ◽  
Takehiko Yoko-o ◽  
Yoh-ichi Shimma ◽  
Yoshifumi Jigami
Keyword(s):  
Cell Wall ◽  
Repetitive Sequence ◽  
Functional Role ◽  
Fluorescent Protein ◽  
Microscopic Analysis ◽  
Red Fluorescent Protein ◽  
Terminal Sequence ◽  
Cell Wall Binding ◽  
Covalently Linked

ABSTRACT We examined the localization of the Pir protein family (Pir1 to Pir4), which is covalently linked to the cell wall in an unknown manner. In contrast to the other Pir proteins, a fusion of Pir1p and monomeric red fluorescent protein distributed in clusters in pir1Δ cells throughout the period of cultivation, indicating that Pir1p is localized in bud scars. Further microscopic analysis revealed that Pir1p is expressed inside the chitin rings of the bud scars. Stepwise deletion of the eight units of the repetitive sequence of Pir1p revealed that one unit is enough for the protein to bind bud scars and that the extent of binding of Pir1p to the cell wall depends on the number of these repetitive units. The localization of a chimeric Pir1p in which the repetitive sequence of Pir1p was replaced with that of Pir4p revealed the functional role of the different protein regions, specifically, that the repetitive sequence is required for binding to the cell wall and that the C-terminal sequence is needed for recruitment to bud scars. This is the first report that bud scars contain proteins like Pir1p as internal components.

A functional role of nuclear polyamines in murine brain

2000 ◽  
Vol 38 ◽  
pp. S66
Author(s):  
K Aramachi
Keyword(s):  
Functional Role ◽  
Murine Brain

Expression and functional role of c-Kit and erbB2 in human hepatoblastoma.

2009 ◽  
Vol 221 (03) ◽  
Author(s):  
B Steiger ◽  
I Leuschner ◽  
D Denkhaus ◽  
D von Schweinitz ◽  
T Pietsch
Keyword(s):  
Functional Role

A functional role of Macrophage Migration Inhibitory Factor (MIF) for liver steatosis in mice by activation of AMP kinase

2012 ◽  
Vol 50 (01) ◽  
Author(s):  
D Heinrichs ◽  
M Knauel ◽  
A Nellen ◽  
P Schmitz ◽  
C Trautwein ◽  
...  
Keyword(s):  
Migration Inhibitory Factor ◽  
Functional Role ◽  
Macrophage Migration Inhibitory Factor ◽  
Liver Steatosis ◽  
Inhibitory Factor ◽  
Macrophage Migration ◽  
Amp Kinase
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