CoChat
Sign Up
Cited by 207Glycosylation and Glycoproteins ResearchViral Infectious Diseases and Gene Expression in InsectsCarbohydrate Chemistry and Synthesis

Glycomics profiling of Chinese hamster ovary cell glycosylation mutants reveals N-glycans of a novel size and complexity

Open access
North, Simon J., Huang, Hung-Hsiang, Sundaram, Subha, Jang-Lee, Jihye, Etienne, A. Tony, Trollope, Alana, Chalabi, Sara, Dell, Anne +2 more·2010·Journal of Biological Chemistry
DOI 10.1074/jbc.m109.068353PMC PMC2820803
Open this paper in CoChat — free accountOpen PDF

Abstract

Identifying biological roles for mammalian glycans and the pathways by which they are synthesized has been greatly facilitated by investigations of glycosylation mutants of cultured cell lines and model organisms. Chinese hamster ovary (CHO) glycosylation mutants isolated on the basis of their lectin resistance have been particularly useful for glycosylation engineering of recombinant glycoproteins. To further enhance the application of these mutants, and to obtain insights into the effects of altering one specific glycosyltransferase or glycosylation activity on the overall expression of cellular glycans, an analysis of the N-glycans and major O-glycans of a panel of CHO mutants was performed using glycomic analyses anchored by matrix-assisted laser desorption ionization-time of flight/time of flight mass spectrometry. We report here the complement of the major N-glycans and O-glycans present in nine distinct CHO glycosylation mutants. Parent CHO cells grown in monolayer versus suspension culture had similar profiles of N- and O-GalNAc glycans, although the profiles of glycosylation mutants Lec1, Lec2, Lec3.2.8.1, Lec4, LEC10, LEC11, LEC12, Lec13, and LEC30 were consistent with available genetic and biochemical data. However, the complexity of the range of N-glycans observed was unexpected. Several of the complex N-glycan profiles contained structures of m/z approximately 13,000 representing complex N-glycans with a total of 26 N-acetyllactosamine (Gal beta1-4GlcNAc)(n) units. Importantly, the LEC11, LEC12, and LEC30 CHO mutants exhibited unique complements of fucosylated complex N-glycans terminating in Lewis(x) and sialyl-Lewis(x) determinants. This analysis reveals the larger-than-expected complexity of N-glycans in CHO cell mutants that may be used in a broad variety of functional glycomics studies and for making recombinant glycoproteins.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1074/jbc.m109.068353

Cites

44 of 44 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Advances in various omics technologies have significantly improved understanding of CHO cells at different levels and enabled rational targeting of glycosylation effectors for improved biopharmaceutical production.

    “Advances in various omics technologies have significantly improved our understanding of CHO cells at different levels and enabled the rational targeting of glycosylation effectors for improved biopharmaceutical production [27,71,72,150–152].”

    From Glycoengineering Chinese hamster ovary cells: a short history (2021) · cites this paper as [27]

    The Introduction (page not provided) says glycomics profiles of nine CHO mutants provide a baseline for future glycosylation engineering, and the Abstract links those profiles to understanding the effects of altered glycosylation activity. The paper does not establish the broader claim about advances across various omics technologies or show that its profiling enabled rational targeting that improved biopharmaceutical production. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • α2,3-linked NeuAc (main sialylated species in CHO-derived glycoproteins) can reduce conformational stability of the CH2 domain of IgG, likely due to a steric effect between the protein backbone and the α2,3-sialylated 6-arm.

    “when NeuAc is α2,3-linked (main sialylated species in CHO-derived glycoproteins) the conformational stability of the CH2 domain of IgG can be reduced, most likely a result of a steric effect between the protein backbone and the α2,3-sialylated 6-arm [27,28].”

    From Glycoengineering Chinese hamster ovary cells: a short history (2021) · cites this paper as [27]

    The Results and Discussion (page unavailable) report that sialic acid in wild-type CHO N-glycans is predominantly NeuAc. The passages do not establish that it is predominantly α2,3-linked, or address IgG CH2-domain stability or the proposed steric effect of a sialylated 6-arm. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • CHO cells do not express alpha-1,3-galactosyltransferases (although they have the gene), and expression of CMP-N-acetylneuraminic acid hydroxylase is virtually absent.

    “CHO cells do not express alpha-1,3-galactosyltransferases, which produce the immunogenic alpha-galactosyl epitope (although they have the gene), and the expression of CMP-N-acetylneuraminic acid hydroxylase, which produces CMP-NeuGc, is virtually absent [27,71,72].”

    From Glycoengineering Chinese hamster ovary cells: a short history (2021) · cites this paper as [27]

    The Results and Discussion section discusses fucosyltransferase genes in CHO cells, but not the α-1,3-galactosyltransferase gene or its expression. It also mentions glycans with terminal NeuGc without addressing CMP-N-acetylneuraminic acid hydroxylase expression. Searches for the named enzymes, the α-galactosyl epitope, and NeuGc did not locate support for either attributed expression claim. (LLM, conf=0.72)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 42

  1. [1]Patnaik S. K. Stanley P. (2006) Methods Enzymol. 416, 159– 182 17113866
  2. [2]Stanley P. (1984) Annu. Rev. Genet. 18, 525– 552 6241454
  3. [3]Narasimhan S. (1982) J. Biol. Chem. 257, 10235– 10242 6213618
  4. [4]Howard D. R. Fukuda M. Fukuda M. N. Stanley P. (1987) J. Biol. Chem. 262, 16830– 16837 2890642
  5. [5]Babu P. North S. J. Jang-Lee J. Chalabi S. Mackerness K. Stowell S. R. Cummings R. D. Rankin S. Dell A. Haslam S. M. (2009) Glycoconj. J. 26, 975– 986 18587645 PMC2791480
  6. [6]Kawar Z. S. Haslam S. M. Morris H. R. Dell A. Cummings R. D. (2005) J. Biol. Chem. 280, 12810– 12819 15653684
  7. [7]Lee J. Park S. H. Sundaram S. Raju T. S. Shaper N. L. Stanley P. (2003) Biochemistry 42, 12349– 12357 14567696
  8. [9]Dell A. Reason A. J. Khoo K. H. Panico M. McDowell R. A. Morris H. R. (1994) Methods Enzymol. 230, 108– 132 8139492
  9. [11]Goldberg D. Sutton-Smith M. Paulson J. Dell A. (2005) Proteomics 5, 865– 875 15693066
  10. [12]Ceroni A. Maass K. Geyer H. Geyer R. Dell A. Haslam S. M. (2008) J. Proteome Res. 7, 1650– 1659 18311910
  11. [13]Wada Y. Azadi P. Costello C. E. Dell A. Dwek R. A. Geyer H. Geyer R. Kakehi K. Karlsson N. G. Kato K. Kawasaki N. Khoo K. H. Kim S. Kondo A. Lattova E. Mechref Y. Miyoshi E. Nakamura K. Narimatsu H. Novotny M. V. Packer N. H. Perreault H. Peter-Katalinic J. Pohlentz G. Reinhold V. N. Rudd P. M. Suzuki A. Taniguchi N. (2007) Glycobiology 17, 411– 422 17223647
  12. [14]Fukuda M. N. Matsumura G. (1976) J. Biol. Chem. 251, 6218– 6225 135762
  13. [15]Goldberg D. Bern M. North S. J. Haslam S. M. Dell A. (2009) Bioinformatics 25, 365– 371 19073587 PMC2732296
  14. [16]Bardor M. Nguyen D. H. Diaz S. Varki A. (2005) J. Biol. Chem. 280, 4228– 4237 15557321
  15. [17]Chen W. Stanley P. (2003) Glycobiology 13, 43– 50 12634323
  16. [18]Varki A. (2008) Trends Mol. Med. 14, 351– 360 18606570 PMC2553044
  17. [19]Stanley P. Sundaram S. Tang J. Shi S. (2005) Glycobiology 15, 43– 53 15329358
  18. [20]Campbell C. Stanley P. (1984) J. Biol. Chem. 259, 13370– 13378 6238035
  19. [21]Sallustio S. Stanley P. (1989) Somat. Cell Mol. Genet. 15, 387– 400 2528829
  20. [22]Potvin B. Stanley P. (1991) Cell Regul. 2, 989– 1000 1724918 PMC361899
  21. [23]Eckhardt M. Gotza B. Gerardy-Schahn R. (1998) J. Biol. Chem. 273, 20189– 20195 9685366
  22. [24]Stanley P. Sudo T. Carver J. P. (1980) J. Cell Biol. 85, 60– 69 7364875 PMC2110602
  23. [25]Lim S. F. Lee M. M. Zhang P. Song Z. (2008) Glycobiology 18, 851– 860 18713811 PMC2722418
  24. [26]Ohyama C. Smith P. L. Angata K. Fukuda M. N. Lowe J. B. Fukuda M. (1998) J. Biol. Chem. 273, 14582– 14587 9603974
  25. [27]Sullivan F. X. Kumar R. Kriz R. Stahl M. Xu G. Y. Rouse J. Chang X. J. Boodhoo A. Potvin B. Cumming D. A. (1998) J. Biol. Chem. 273, 8193– 8202 9525924
  26. [28]Ripka J. Adamany A. Stanley P. (1986) Arch. Biochem. Biophys. 249, 533– 545 2428310
  27. [29]Becker D. J. Lowe J. B. (2003) Glycobiology 13, 41R– 53R 12651883
  28. [30]Chen W. Tang J. Stanley P. (2005) Glycobiology 15, 259– 269 15525819
  29. [31]Kanda Y. Yamane-Ohnuki N. Sakai N. Yamano K. Nakano R. Inoue M. Misaka H. Iida S. Wakitani M. Konno Y. Yano K. Shitara K. Hosoi S. Satoh M. (2006) Biotechnol. Bioeng. 94, 680– 688 16609957
  30. [32]Moloney D. J. Shair L. H. Lu F. M. Xia J. Locke R. Matta K. L. Haltiwanger R. S. (2000) J. Biol. Chem. 275, 9604– 9611 10734111
  31. [33]Weinstein J. Sundaram S. Wang X. Delgado D. Basu R. Stanley P. (1996) J. Biol. Chem. 271, 27462– 27469 8910328
  32. [34]Zhao Y. Sato Y. Isaji T. Fukuda T. Matsumoto A. Miyoshi E. Gu J. Taniguchi N. (2008) FEBS J. 275, 1939– 1948 18384383
  33. [35]Hong Y. Stanley P. (2003) J. Biol. Chem. 278, 53045– 53054 14561743
  34. [36]Stanley P. (1989) Mol. Cell. Biol. 9, 377– 383 2710109 PMC362611
  35. [37]Butters T. D. Sparks L. M. Harlos K. Ikemizu S. Stuart D. I. Jones E. Y. Davis S. J. (1999) Protein Sci. 8, 1696– 1701 10452614 PMC2144425
  36. [38]Wang J. H. Smolyar A. Tan K. Liu J. H. Kim M. Sun Z. Y. Wagner G. Reinherz E. L. (1999) Cell 97, 791– 803 10380930
  37. [39]Zhang A. Potvin B. Zaiman A. Chen W. Kumar R. Phillips L. Stanley P. (1999) J. Biol. Chem. 274, 10439– 10450 10187834
  38. [40]Patnaik S. K. Zhang A. Shi S. Stanley P. (2000) Arch. Biochem. Biophys. 375, 322– 332 10700388
  39. [41]Nishihara S. Iwasaki H. Kaneko M. Tawada A. Ito M. Narimatsu H. (1999) FEBS Lett. 462, 289– 294 10622713
  40. [42]Raman R. Raguram S. Venkataraman G. Paulson J. C. Sasisekharan R. (2005) Nat. Methods 2, 817– 824 16278650
  41. [43]Raman R. Venkataraman M. Ramakrishnan S. Lang W. Raguram S. Sasisekharan R. (2006) Glycobiology 16, 82R– 90R 16478800
  42. [44]Solter D. Knowles B. B. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 5565– 5569 281705 PMC393007

Cited by

Sorted by citations

Related

Authors
North, S. J., Huang, H.-H., Sundaram, S., Jang-Lee, J., Etienne, A. T., Trollope, A., Chalabi, S., Dell, A., Stanley, P., & Haslam, S. M.
Affiliations
  • Imperial College London (GB)
  • Albert Einstein College of Medicine (US)
Corresponding author
  • Stanley, Pamela
  • Haslam, Stuart M.
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2010-02
Volume
285
Issue
8
Pages
5759-5775
ISSN
0021-9258 (ISSN-L)
DOI
10.1074/jbc.m109.068353
PMC
PMC2820803
OpenAlex
W2033604747
Cited by
207
References
44
Citations by year
2026: 5 · 2025: 7 · 2024: 8 · 2023: 6 · 2022: 10 · 2021: 12 · 2020: 11 · 2019: 15 · 2018: 17 · 2017: 16 · 2016: 8 · 2015: 11 · 2014: 21 · 2013: 17 · 2012: 13
FWCI
5.78
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
Funders
  • Wellcome Trust
  • National Institutes of Health
  • Directorate for Biological Sciences
  • U.S. Public Health Service (NCI RO1-36434)
  • Biotechnology and Biological Sciences Research Council (BB/F008309/1, B19088)
  • National Cancer Institute
  • National Institute of General Medical Sciences (GM 62116, U54GM062116)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
Keywords
glycosylation mutants, CHO cells, N-glycans, O-glycans, glycomics, lectin resistance, glycosyltransferases, N-acetyllactosamine, fucosylated N-glycans, CD15, sialyl Lewis X
MeSH terms
Animals, Carbohydrate Sequence, Cricetulus, Glycosylation, Cricetinae, Mutation, Polysaccharides, CHO Cells

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review