title: Digital mRNA profiling of N-glycosylation gene expression in recombinant Chinese hamster ovary cells treated with sodium butyrate.
abstract: To understand the effects of sodium butyrate (NaBu) on protein glycosylation, recombinant Chinese hamster ovary (rCHO) cells producing Fc-fusion glycoprotein were subjected to 3mM NaBu. The addition of NaBu to the cultures reduced the relative proportion of acidic isoforms and sialic acid content of the glycoprotein. Fifty-two N-glycosylation-related gene expressions were also assessed by the NanoString nCounter system, which can provide a direct digital readout using custom-designed color-coded probes. Among them, ten genes (ugp, slc35a2, ganc, man1a, man1c, mgat5a, st3gal5, glb1, neu1, and neu3) were up-regulated and three genes (b4galt2, st3gal3, and neu2) were down-regulated significantly. Altered expression patterns in st3gal3, neu1, and neu3, which have roles in the sialic acid biosynthesis pathway, correlated with reduced sialic acid content of the glycoprotein by NaBu. Taken together, the results obtained in this study provide a better understanding of the detrimental effect of NaBu on N-glycosylation in rCHO cells.
authors: Department of Biological Sciences, KAIST, 373-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea.
authors: Sang Min
authors: SM
authors: Lee
authors: Biotechnology Process Engineering Center, KRIBB, Daejon 305-806, Korea.
authors: Yeon-Gu
authors: YG
authors: Kim
authors: Biotechnology Process Engineering Center, KRIBB, Daejon 305-806, Korea. Electronic address: eglee@kribb.re.kr.
authors: Eun Gyo
authors: EG
authors: Lee
authors: Department of Biological Sciences, KAIST, 373-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea. Electronic address: gyunminlee@kaist.ac.kr.
authors: Gyun Min
authors: GM
authors: Lee
mesh_terms: D000818:Animals; D020148:Butyric Acid / Q000494:pharmacology*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D020869:Gene Expression Profiling / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D006031:Glycosylation; D007525:Isoelectric Focusing; D014176:Protein Biosynthesis / Q000187:drug effects* / Q000235:genetics; D011499:Protein Processing, Post-Translational / Q000187:drug effects* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D020148:Butyric Acid
keywords: CHO cells; Gene expression; N-Glycosylation; NanoString nCounter system; Sodium butyrate