Inhibition of Nucleotide Sugar Transport in Trypanosoma brucei Alters Surface Glycosylation

Liu, Li; Xu, Yuxin; Caradonna, Kacey L.; Kruzel, Emilia K.; Burleigh, Barbara A.; Bangs, James D.; Hirschberg, Carlos B.

doi:10.1074/jbc.m113.453597

Cited by 15 publications

(13 citation statements)

References 78 publications

(96 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To identify UDP-GlcNAc transporters, we used a yeast complementation approach in which a Kluyveromyces lactis mutant deficient in the Golgi transport of UDP-GlcNAc was used [ 19 ]. This mutant has been used for the identification of UDP-GlcNAc transporters from canine cells [ 20 ], Caenorhabditis elegans [ 21 ] and T. brucei [ 13 ]. Complementation assays are based on the less intense binding of the GS-II lectin from Griffonia simplicifolia , which specifically recognizes terminal GlcNAc residues, to the mutant cell surface.…”

Section: Resultsmentioning

confidence: 99%

“…In parasitic protozoa, the role of NSTs has been investigated in Leishmania spp. [ 11 , 12 ], T. brucei [ 13 ] and Toxoplasma gondi [ 14 ]. In L. major , knockout of the GDP-mannose (GDP-Man) transporter LPG2 resulted in mutants deficient in cell-surface phosphoglycan molecules displaying attenuated virulence in mice [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

“…In L. major , knockout of the GDP-mannose (GDP-Man) transporter LPG2 resulted in mutants deficient in cell-surface phosphoglycan molecules displaying attenuated virulence in mice [ 15 ]. Recently, four NSTs from T. brucei (TbNST1-4) were characterized [ 13 ]. Silencing and knockout experiments of TbNST4, which transports UDP- N -acetylglucosamine (UDP-GlcNAc), UDP- N -acetylgalactosamine (UDP-GalNAc) and GDP-Man, resulted in glycosylation defects, but no impairment of infectivity was observed, likely due to redundancy in function.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Identification of a Golgi-localized UDP-N-acetylglucosamine transporter in Trypanosoma cruzi

et al. 2015

View full text Add to dashboard Cite

BackgroundNucleotide sugar transporters (NSTs) play an essential role in translocating nucleotide sugars into the lumen of the endoplasmic reticulum and Golgi apparatus to be used as substrates in glycosylation reactions. This intracellular transport is an essential step in the biosynthesis of glycoconjugates.ResultsWe have identified a family of 11 putative NSTs in Trypanosoma cruzi, the etiological agent of Chagas’ disease. A UDP-N-acetylglucosamine transporter, TcNST1, was identified by a yeast complementation approach. Based on a phylogenetic analysis four candidate genes were selected and used for complementation assays in a Kluyveromyces lactis mutant strain. The transporter is likely expressed in all stages of the parasite life cycle and during differentiation of epimastigotes to infective metacyclics. Immunofluorescence analyses of a GFP-TcNST1 fusion protein indicate that the transporter is localized to the Golgi apparatus. As many NSTs are multisubstrate transporters, we also tested the capacity of TcNST1 to transport GDP-Man.ConclusionsWe have identified a UDP-N-acetylglucosamine transporter in T. cruzi, which is specifically localized to the Golgi apparatus and seems to be expressed, at the mRNA level, throughout the parasite life cycle. Functional studies of TcNST1 will be important to unravel the role of NSTs and specific glycoconjugates in T. cruzi survival and infectivity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0601-7) contains supplementary material, which is available to authorized users.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Identification of a Golgi-localized UDP-N-acetylglucosamine transporter in Trypanosoma cruzi

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Studies that showed the occurrence of nucleotide sugar transporters in protozoan parasites established additional new concepts. In collaboration with John Samuelson's group, Luis Bredeston of my lab found endoplasmic reticulum and Golgi apparatus functions in these organisms (25), and in collaboration with Jay Bangs and Barbara Burleigh's groups, Li Liu and Yu Xin Xu of my lab described a nucleotide sugar transporter that transports both purine and pyrimidine substrates in Trypanosoma brucei, something we had never seen in any organism before (26). Together with John Boothroyd's group, Carolina Caffaro showed that a deletion of a transporter gene in Toxoplasma gondii contributed to changes in infectivity (27).…”

Section: Boston Mamentioning

confidence: 99%

My journey in the discovery of nucleotide sugar transporters of the Golgi apparatus

Hirschberg

2018

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Defects in protein glycosylation can have a dramatic impact on eukaryotic cells and is associated with mental and developmental pathologies in humans. The studies outlined below illustrate how a basic biochemical problem in the mechanisms of protein glycosylation, specifically substrate transporters of nucleotide sugars, including ATP and 3'-phosphoadenyl-5'-phosphosulfate (PAPS), in the membrane of the Golgi apparatus and endoplasmic reticulum, expanded into diverse biological systems from mammals, including humans, to yeast, roundworms, and protozoa. Using these diverse model systems allowed my colleagues and me to answer fundamental biological questions that enabled us to formulate far-reaching hypotheses and expanded our knowledge of human diseases caused by malfunctions in the metabolic processes involved.

show abstract

“…Ammonium plays an active part in the regulation of the pH of acidic intracellular compartments of CHO expressing recombinant immunoadhesin tumor necrosis factor-IgG (TNFR-IgG), including trans-Golgi and it affects intracellular nucleotide sugar pools, especially UDP-N-acetylglucosamine (UDP-GlcNAc) and UDP-N-acetylgalactosamine 237 . The availability of nucleotide sugars in the lumen of the Golgi apparatus also depends on the transport, as shown in Trypanosoma brucei, where the inhibition of transporters modified surface glycosylation 286 . Increasing levels of glycosylation precursors, as result of cell culture medium supplementation for instance, alters the glycan distribution 124 .…”

Section: Introductionmentioning

confidence: 99%

Tailoring recombinant protein quality by rational media design

Brühlmann

Jordan

Hemberger³

et al. 2015

Biotechnology Progress

View full text Add to dashboard Cite

Clinical efficacy and safety of recombinant proteins are closely associated with their structural characteristics. The major quality attributes comprise glycosylation, charge variants (oxidation, deamidation, and C‐ & N‐terminal modifications), aggregates, low‐molecular‐weight species (LMW), and misincorporation of amino acids in the protein backbone. Cell culture media design has a great potential to modulate these quality attributes due to the vital role of medium in mammalian cell culture. The purpose of this review is to provide an overview of the way both classical cell culture medium components and novel supplements affect the quality attributes of recombinant therapeutic proteins expressed in mammalian hosts, allowing rational and high‐throughput optimization of mammalian cell culture media. A selection of specific and/or potent inhibitors and activators of oligosaccharide processing as well as components affecting multiple quality attributes are presented. Extensive research efforts in this field show the feasibility of quality engineering through media design, allowing to significantly modulate the protein function. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:615–629, 2015

show abstract

Inhibition of Nucleotide Sugar Transport in Trypanosoma brucei Alters Surface Glycosylation

Cited by 15 publications

References 78 publications

Identification of a Golgi-localized UDP-N-acetylglucosamine transporter in Trypanosoma cruzi

Identification of a Golgi-localized UDP-N-acetylglucosamine transporter in Trypanosoma cruzi

My journey in the discovery of nucleotide sugar transporters of the Golgi apparatus

Tailoring recombinant protein quality by rational media design

Contact Info

Product

Resources

About