Pemprofilan KCPT pendarfluor dan penyerakan cahaya sejatan oligosakarida terpaut-(N) asparagina intrasel daripada saccharomyces cerevisiae menggunakan mutan alg8

Translated title of the contribution: Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant

Iqbal Jalaludin, Amirul Husna Sudin, Ikram Mohd Said, Kamalrul Azlan Azizan, Syarul Nataqain Baharum, Abdul Munir Abd. Murad, Farah Diba Abu Bakar, Nor Muhammad Mahadi, Mark R. Wormald, Dominic S. Alonzi, Muhammad Mukram Mohamed Mackeen

Research output: Contribution to journalArticle

Abstract

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.

Original languageMalay
Pages (from-to)1210-1218
Number of pages9
JournalMalaysian Journal of Analytical Sciences
Volume21
Issue number6
DOIs
Publication statusPublished - 1 Dec 2017

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Asparagine
Oligosaccharides
Light scattering
Yeast
Fluorescence
Glycosylation
Glycoproteins
Polysaccharides
Bacteria
Genes
lipid-linked oligosaccharides
Proteins

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

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title = "Pemprofilan KCPT pendarfluor dan penyerakan cahaya sejatan oligosakarida terpaut-(N) asparagina intrasel daripada saccharomyces cerevisiae menggunakan mutan alg8",
abstract = "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.",
keywords = "Free oligosaccharides, Lipid-linked oligosaccharides, N-glycans, N-linked glycosylation, Saccharomyces cerevisiae",
author = "Iqbal Jalaludin and Sudin, {Amirul Husna} and Said, {Ikram Mohd} and Azizan, {Kamalrul Azlan} and Baharum, {Syarul Nataqain} and {Abd. Murad}, {Abdul Munir} and {Abu Bakar}, {Farah Diba} and Mahadi, {Nor Muhammad} and Wormald, {Mark R.} and Alonzi, {Dominic S.} and {Mohamed Mackeen}, {Muhammad Mukram}",
year = "2017",
month = "12",
day = "1",
doi = "10.17576/mjas-2017-2106-01",
language = "Malay",
volume = "21",
pages = "1210--1218",
journal = "Malaysian Journal of Analytical Sciences",
issn = "1394-2506",
publisher = "Faculty of Science and Technology, Universiti Kebangsaan Malaysia",
number = "6",

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TY - JOUR

T1 - Pemprofilan KCPT pendarfluor dan penyerakan cahaya sejatan oligosakarida terpaut-(N) asparagina intrasel daripada saccharomyces cerevisiae menggunakan mutan alg8

AU - Jalaludin, Iqbal

AU - Sudin, Amirul Husna

AU - Said, Ikram Mohd

AU - Azizan, Kamalrul Azlan

AU - Baharum, Syarul Nataqain

AU - Abd. Murad, Abdul Munir

AU - Abu Bakar, Farah Diba

AU - Mahadi, Nor Muhammad

AU - Wormald, Mark R.

AU - Alonzi, Dominic S.

AU - Mohamed Mackeen, Muhammad Mukram

PY - 2017/12/1

Y1 - 2017/12/1

N2 - 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.

AB - 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.

KW - Free oligosaccharides

KW - Lipid-linked oligosaccharides

KW - N-glycans

KW - N-linked glycosylation

KW - Saccharomyces cerevisiae

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U2 - 10.17576/mjas-2017-2106-01

DO - 10.17576/mjas-2017-2106-01

M3 - Article

VL - 21

SP - 1210

EP - 1218

JO - Malaysian Journal of Analytical Sciences

JF - Malaysian Journal of Analytical Sciences

SN - 1394-2506

IS - 6

ER -