Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant
N-glycans are biologically important oligosaccharides associated with the asparagine residue that may exist in protein-bound or unbound forms in all eukaryotes (including yeasts) and some bacteria. The- core structure of these oligosaccharides is based on the trimannosyl chitobiose structure result...
| Main Authors: | , , , , , , , , , , |
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| Format: | Journal article |
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Malaysian Analytical Sciences Society
2017
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| _version_ | 1797105022355374080 |
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| author | Jalaludin, I Sudin, AH Said, IM Azizan, KA Baharum, SN Murad, AMA Bakar, FDA Mahadi, NM Wormald, MR Alonzi, DS Mackeen, MM |
| author_facet | Jalaludin, I Sudin, AH Said, IM Azizan, KA Baharum, SN Murad, AMA Bakar, FDA Mahadi, NM Wormald, MR Alonzi, DS Mackeen, MM |
| author_sort | Jalaludin, I |
| collection | OXFORD |
| description | N-glycans are biologically important oligosaccharides associated with the asparagine residue that may exist in protein-bound or unbound forms in all eukaryotes (including yeasts) and some bacteria. The- core structure of these oligosaccharides is based on the trimannosyl chitobiose structure resulting from cellular N-glycosylation. Preparative-scale amounts of these oligosaccharides are important for chemical, structural and functional studies due to their biological significance. Therefore, we explored a biochemical approach of oligosaccharide preparation using mutant-derived monoglucosylated lipid-linked oligosaccharides (LLOs) required for the assembly of N-linked glycoproteins and non-monoglucosylated free-oligosaccharides (fOSs) from misfolded N-linked glycoproteins using an N-glycosylation (alg) mutant of Saccharomyces cerevisiae. Oligosaccharide extracts of fOSs and LLOs from the alg8 S. cerevisiae mutant lacking the ALG8 gene were profiled using fluorescence- and evaporative light scattering-based HPLC. LLOs did not produce accumulated levels of the target mutant- related monoglucosylated (Glc1Man9GlcNAc2) at 100 ml scale. However, it was possible to detect truncated oligomannose (paucimannose) structures in the fOSs of the alg8 mutant. |
| first_indexed | 2024-03-07T06:41:36Z |
| format | Journal article |
| id | oxford-uuid:f9772cf4-cec0-45f7-8a69-858ce9e60466 |
| institution | University of Oxford |
| last_indexed | 2024-03-07T06:41:36Z |
| publishDate | 2017 |
| publisher | Malaysian Analytical Sciences Society |
| record_format | dspace |
| spelling | oxford-uuid:f9772cf4-cec0-45f7-8a69-858ce9e604662022-03-27T12:58:11ZFluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutantJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:f9772cf4-cec0-45f7-8a69-858ce9e60466Symplectic Elements at OxfordMalaysian Analytical Sciences Society2017Jalaludin, ISudin, AHSaid, IMAzizan, KABaharum, SNMurad, AMABakar, FDAMahadi, NMWormald, MRAlonzi, DSMackeen, MMN-glycans are biologically important oligosaccharides associated with the asparagine residue that may exist in protein-bound or unbound forms in all eukaryotes (including yeasts) and some bacteria. The- core structure of these oligosaccharides is based on the trimannosyl chitobiose structure resulting from cellular N-glycosylation. Preparative-scale amounts of these oligosaccharides are important for chemical, structural and functional studies due to their biological significance. Therefore, we explored a biochemical approach of oligosaccharide preparation using mutant-derived monoglucosylated lipid-linked oligosaccharides (LLOs) required for the assembly of N-linked glycoproteins and non-monoglucosylated free-oligosaccharides (fOSs) from misfolded N-linked glycoproteins using an N-glycosylation (alg) mutant of Saccharomyces cerevisiae. Oligosaccharide extracts of fOSs and LLOs from the alg8 S. cerevisiae mutant lacking the ALG8 gene were profiled using fluorescence- and evaporative light scattering-based HPLC. LLOs did not produce accumulated levels of the target mutant- related monoglucosylated (Glc1Man9GlcNAc2) at 100 ml scale. However, it was possible to detect truncated oligomannose (paucimannose) structures in the fOSs of the alg8 mutant. |
| spellingShingle | Jalaludin, I Sudin, AH Said, IM Azizan, KA Baharum, SN Murad, AMA Bakar, FDA Mahadi, NM Wormald, MR Alonzi, DS Mackeen, MM Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant |
| title | Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant |
| title_full | Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant |
| title_fullStr | Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant |
| title_full_unstemmed | Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant |
| title_short | Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant |
| title_sort | fluorescence and evaporative light scattering hplc profiling of intracellular asparagine n linked oligosaccharides from saccharomyces cerevisiae using the alg8 mutant |
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