Sodium butyrate alters erythropoietin glycosylation via multiple mechanisms
| Autor: | Qiang Qin, Christopher K. Crowell, Gustavo Grampp, Richard A. Radcliffe, Robert I. Scheinman, Gary Rogers |
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| Rok vydání: | 2007 |
| Předmět: |
Glycan
Glycosylation Fibrosarcoma Bioengineering Biology Protein Engineering Applied Microbiology and Biotechnology Models Biological Glycomics chemistry.chemical_compound Cell Line Tumor Humans Computer Simulation skin and connective tissue diseases Gene Erythropoietin chemistry.chemical_classification Dose-Response Relationship Drug Sodium butyrate Molecular biology Recombinant Proteins Neoplasm Proteins carbohydrates (lipids) Gene Expression Regulation Neoplastic Butyrates chemistry Biochemistry Cell culture biology.protein HT1080 sense organs Glycoprotein Biotechnology |
| Zdroj: | Biotechnology and bioengineering. 99(1) |
| ISSN: | 1097-0290 |
| Popis: | Recombinant human erythropoietin (rHuEPO) produced in a human kidney fibrosarcoma cell line, HT1080, was used as a model to study the effects of sodium butyrate (SB) on protein glycosylation. Treatment with 2 mM SB resulted in complex changes with respect to sugar nucleotide pools including an increase in UDP-Gal and a decrease in UDP-GlcNac. In addition, polylactosamine structures present on rHuEPO increased after SB treatment. To determine if these phenotypic changes correlated with changes in mRNA abundance, we profiled mRNA levels over a 24-h period in the presence or absence of SB using oligonucleotide microarrays. By filtering our data through a functional glycomics gene list associated with the processes of glycan degradation, glycan synthesis, and sugar nucleotide synthesis and transport we identified 26 genes with significantly altered mRNA levels. We were able to correlate the changes in message in six of these genes with measurable phenotypic changes within our system including: neu1, b3gnt6, siat4b, b3gnt1, slc17a5, and galt. Interestingly, for the two genes: cmas and gale, our measurable phenotypic changes did not correlate with changes in mRNA expression. These data demonstrate both the utility and pit falls of coupling biochemical analysis with high throughput oligonucleotide microarrays to predict how changes in cell culture environments will impact glycoprotein oligosaccharide content. |
| Databáze: | OpenAIRE |
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