The effect of ammonia on the O-linked glycosylation of granulocyte colony-stimulating factor produced by chinese hamster ovary cells

Andersen, D C; Goochee, C F

doi:10.1002/bit.260470112

Title: The effect of ammonia on the O-linked glycosylation of granulocyte colony-stimulating factor produced by chinese hamster ovary cells

Full Record
Other Related Research

Abstract

Ammonium ion concentrations ranging from 0 to 10 mM are shown to significantly reduce the sialylation of granulocyte colony-stimulating factor (G-CSF) produced by recombinant Chinese hamster ovary cells. Specifically, the degree of completion of the final reaction in the O-linked glycosylation pathway, the addition of sialic acid in an {alpha}(2,6) linkage to N-acetylgalactosamine, is reduced by NH{sub 4}{sup +} concentrations of as low as 2 mM. The effect of ammonia on sialylation is rapid, sustained, and does not affect the secretion rate of G-CSF. Additionally, the effect can be mimicked using the weak base chloroquine, suggesting that the effect is related to the weak base characteristics of ammonia. In support of this hypothesis, experiments using brefeldin A suggest that the addition of sialic acid in an {alpha}(2,6) linkage to N-acetylgalactosamine occurs in the trans-Golgi compartment prior to the trans-Golgi network, which would be expected under normal conditions to have a slightly acidic pH in the range from 6.5 to 6.75. Ammonium ion concentrations of 10mM would be expected to reduce significantly the differences in pH between acidic intracellular compartments and the cytoplasm. The pH-activity profile for the CHO O-linked {alpha}(2,6)sialytransferase using monosialylated G-CSF as a substrate reveals a twofold decreasemore »« less

Authors:

Andersen, D C; Goochee, C F ^[1]

Stanford Univ., CA (United States). Dept. of Chemical Engineering

Publication Date:: Wed Jul 05 00:00:00 EDT 1995

Sponsoring Org.:: USDOE

OSTI Identifier:: 78139

Resource Type:: Journal Article

Journal Name:: Biotechnology and Bioengineering

Additional Journal Information:: Journal Volume: 47; Journal Issue: 1; Other Information: PBD: 5 Jul 1995

Country of Publication:: United States

Language:: English

Subject:: 56 BIOLOGY AND MEDICINE, APPLIED STUDIES; AMMONIA; BIOLOGICAL EFFECTS; CHO CELLS; SENSITIVITY; GROWTH FACTORS; METABOLISM; BIOLOGICAL PATHWAYS; TRANSFERASES

Citation Formats


                    Andersen, D C, and Goochee, C F. The effect of ammonia on the O-linked glycosylation of granulocyte colony-stimulating factor produced by chinese hamster ovary cells.  United States: N. p., 1995. 
        Web.  doi:10.1002/bit.260470112.

Copy to clipboard


                    Andersen, D C, & Goochee, C F. The effect of ammonia on the O-linked glycosylation of granulocyte colony-stimulating factor produced by chinese hamster ovary cells.  United States.  https://doi.org/10.1002/bit.260470112

Copy to clipboard


                    Andersen, D C, and Goochee, C F. 1995.  
        "The effect of ammonia on the O-linked glycosylation of granulocyte colony-stimulating factor produced by chinese hamster ovary cells".  United States.  https://doi.org/10.1002/bit.260470112.

Copy to clipboard


                    
@article{osti_78139,

  title        = {The effect of ammonia on the O-linked glycosylation of granulocyte colony-stimulating factor produced by chinese hamster ovary cells},

  author       = {Andersen, D C and Goochee, C F},

  abstractNote = {Ammonium ion concentrations ranging from 0 to 10 mM are shown to significantly reduce the sialylation of granulocyte colony-stimulating factor (G-CSF) produced by recombinant Chinese hamster ovary cells. Specifically, the degree of completion of the final reaction in the O-linked glycosylation pathway, the addition of sialic acid in an {alpha}(2,6) linkage to N-acetylgalactosamine, is reduced by NH{sub 4}{sup +} concentrations of as low as 2 mM. The effect of ammonia on sialylation is rapid, sustained, and does not affect the secretion rate of G-CSF. Additionally, the effect can be mimicked using the weak base chloroquine, suggesting that the effect is related to the weak base characteristics of ammonia. In support of this hypothesis, experiments using brefeldin A suggest that the addition of sialic acid in an {alpha}(2,6) linkage to N-acetylgalactosamine occurs in the trans-Golgi compartment prior to the trans-Golgi network, which would be expected under normal conditions to have a slightly acidic pH in the range from 6.5 to 6.75. Ammonium ion concentrations of 10mM would be expected to reduce significantly the differences in pH between acidic intracellular compartments and the cytoplasm. The pH-activity profile for the CHO O-linked {alpha}(2,6)sialytransferase using monosialylated G-CSF as a substrate reveals a twofold decrease in enzymatic activity across the pH range from 6.75 to 7.0. Mathematical modeling of this sialylation reaction supports the hypothesis that this twofold decrease in sialyltransferase activity resulting from an ammonia-induced increase in trans-Golgi pH could produce the observed decrease in G-CSF sialylation.},

  doi          = {10.1002/bit.260470112},

  url          = {https://www.osti.gov/biblio/78139},
  journal      = {Biotechnology and Bioengineering},
number       = 1,

  volume       = 47,

  place        = {United States},

  year         = {Wed Jul 05 00:00:00 EDT 1995},

  month        = {Wed Jul 05 00:00:00 EDT 1995}

}

Copy to clipboard

Journal Article:

https://doi.org/10.1002/bit.260470112

Other availability

Find in Google Scholar

Search WorldCat to find libraries that may hold this journal

Save / Share:

Export Metadata

Save to My Library

Similar records in OSTI.GOV collections:

Separation of anionic oligosaccharides by high-performance liquid chromatography

Journal Article Green, E; Baenziger, J - Anal. Biochem.; (United States)

The authors have developed methods for rapid fractionation of anionic oligosaccharides containing sulfate and/or sialic acid moieties by high-performance liquid chromatography (HPLC). Ion-exchange HPLC on amine-bearing columns (Micropak AX-10 and AX-5) at pH 4.0 is utilized to separate anionic oligosaccharides bearing zero, one, two, three, or four charges, independent of the identity of the anionic moieties (sulfate and/or sialic acid). Ion-exchange HPLC at pH 1.7 allows separation of neutral, mono-, di-, and tetrasialylated, monosulfated, and disulfated oligosaccharides. Oligosaccharides containing three sialic acid residues and those bearing one each of sulfate and sialic acid, however, coelute at pH 1.7. Since themore »« less
https://doi.org/10.1016/0003-2697(86)90585-3
Asparagine-linked oligosaccharides on lutropin, follitropin, and thyrotropin: distributions of sulfated and sialylated oligosaccharides on bovine, ovine, and human pituitary glycoprotein hormones

Journal Article Green, E; Baenziger, J - J. Biol. Chem.; (United States)

The asparagine-linked oligosaccharides on the pituitary glycoprotein hormones lutropin (LH), follitropin (FSH), and thyrotropin (TSH) consist of a heterogeneous array of neutral, sulfated, sialylated, and sulfated/sialylated structures. In this study, the authors determined the relative quantities of the various asparagine-linked oligosaccharides on LH, FSH, and TSH from these three animal species. The proportions of sulfated versus sialylated oligosaccharides varied markedly among the different hormones. Both hormone- and animal species-specific differences in the types and distributions of sulfated, sialylated, and sulfated/sialylated structures were evident. In particular, LH and FSH, which are synthesized in the same pituitary cell and bear ..cap alpha..-subunitsmore »« less
Purification and characterization of a CMP-sialic:LcOse4Cer sialyltransferase from human colorectal carcinoma cell membranes

Journal Article Liepkans, V; Jolif, A; Larson, G - Biochemistry; (United States)

Purified lactotetraosylceramide (Gal beta 1----3GlcNAc beta 1----3Gal beta 1----4Glc1-Cer) was tested for its ability to accept (14C)sialic acid from CMP-(14C)sialic into monosialoganglioside fractions in the presence of membrane fractions purified from human colorectal carcinoma cells (SW1116). Membrane fractions were isolated by three different methods: sucrose density centrifugation, CMP-agarose gel column chromatography, and LcOse4 gel chromatography. We optimized the incubation conditions for detergent dependency (taurocholate), pH (6.3), and acceptor concentration. The sialyltransferase activity was dependent on membrane protein and linear for time up to at least 4 h. The LcOse4 affinity chromatography of the crude microsomal membrane pellet from these cellsmore »« less
https://doi.org/10.1021/bi00423a026
Subcellular distribution of swine vesicular disease virus proteins and alterations induced in infected cells: A comparative study with foot-and-mouth disease virus and vesicular stomatitis virus

Journal Article Martin-Acebes, Miguel; Gonzalez-Magaldi, Monica; Rosas, Maria; ... - Virology

The intracellular distribution of swine vesicular disease virus (SVDV) proteins and the induced reorganization of endomembranes in IBRS-2 cells were analyzed. Fluorescence to new SVDV capsids appeared first upon infection, concentrated in perinuclear circular structures and colocalized to dsRNA. As in foot-and-mouth disease virus (FMDV)-infected cells, a vesicular pattern was predominantly found in later stages of SVDV capsid morphogenesis that colocalized with those of non-structural proteins 2C, 2BC and 3A. These results suggest that assembly of capsid proteins is associated to the replication complex. Confocal microscopy showed a decreased fluorescence to ER markers (calreticulin and protein disulfide isomerase), and disorganizationmore »« less
https://doi.org/10.1016/j.virol.2007.12.042
Localization of UDP-GlcNAc 2-epimerase/ManAc kinase (GNE) in the Golgi complex and the nucleus of mammalian cells

Journal Article Krause, Sabine; Hinderlich, Stephan; Amsili, Shira; ... - Experimental Cell Research

The bifunctional enzyme UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE) is essential for early embryonic development and catalyzes the rate limiting step in sialic acid biosynthesis. Although epimerase and kinase activities have been attributed to GNE, little is known about the regulation, differential expression, and subcellular localization of GNE in vivo. Mutations in GNE cause a rare inherited muscle disorder in humans called hereditary inclusion body myopathy (HIBM). However, the role of GNE in HIBM pathogenesis has not been defined yet. Here, we show that the GNE protein is expressed in various mammalian cells and tissues with highest levels found in cancer cells andmore »« less
https://doi.org/10.1016/j.yexcr.2004.11.010

Similar Records