Structure of glycosylated Cu/Zn-superoxide dismutase from Kluyveromyces yeast NBIMCC 1984

Dolashka-Angelova, Pavlina; Moshtanska, Vesela; Kujumdzieva, Anna; Атанасов, Б.; Petrova, Ventsislava Yankova; Voelter, Wolfgang; Beeumen, Jozef Van

doi:10.1016/j.molstruc.2010.06.031

Cited by 5 publications

(3 citation statements)

References 38 publications

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“…EC-SOD is known to be the only glycosylated SOD isoenzyme in mammals [5,10], whereas naturally glycosylated Cu,Zn-SOD is known to be present in fungi [11,12] and yeast [13]. A specific biological function of the N-glycan on EC-SOD has not been clearly identified yet [14].…”

Section: Introductionmentioning

confidence: 99%

N-Glycosylation profiling of recombinant mouse extracellular superoxide dismutase produced in Chinese hamster ovary cells

Korekane

Yamaguchi

et al. 2011

Glycoconj J

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Extracellular superoxide dismutase (EC-SOD), the major SOD isoenzyme in biological fluids, is known to be N-glycosylated and heterogeneous as was detected in most glycoproteins. However, only one N-glycan structure has been reported in recombinant human EC-SOD produced in Chinese hamster ovary (CHO) cells. Thus, a precise N-glycan profile of the recombinant EC-SOD is not available. In this study, we report profiling of the N-glycan in the recombinant mouse EC-SOD produced in CHO cells using high-resolution techniques, including the liberation of N-glycans by treatment with PNGase F, fluorescence labeling by pyridylamination, characterization by anion-exchange, normal and reversed phase-HPLC separation, and mass spectrometry. We succeeded in identifying 26 different types of N-glycans in the recombinant enzyme. The EC-SOD N-glycans were basically core-fucosylated (98.3% of the total N-glycan content), and were high mannose sugar chain, and mono-, bi-, tri-, and tetra-antennary complex sugar chains exhibiting varying degrees of sialylation. Four of the identified N-glycans were uniquely modified with a sulfate group, a Lewis(x) structure, or an α-Gal epitope. The findings will shed new light on the structure-function relationships of EC-SOD N-glycans.

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Section: Introductionmentioning

confidence: 99%

N-Glycosylation profiling of recombinant mouse extracellular superoxide dismutase produced in Chinese hamster ovary cells

Korekane

Yamaguchi

et al. 2011

Glycoconj J

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“…Therefore, there is still a challenge to produce a large body of humanized therapeutic end-products with complex glycan in suitable yeasts. Making full use of the CRISPR-Cas9 technique may be a potential strategy to address the important issues (Ueda et al, 1993 ; Dolashka-Angelova et al, 2010 ).…”

Section: Discussionmentioning

confidence: 99%

“…Much effort has been paid to improving the N-glycosylation pathway of P. pastoris to mimic the human N-glycosylation pathway. Successful utilization of the CRISPR-Cas9 system has been used for the humanization of the glycosylation pathways in P. pastoris by disrupting och1 and alg3 (mannosyltransferase) genes (Dolashka-Angelova et al, 2010 ). Kluyveromyces lactis is a food-grade yeast commonly used for industrial applications.…”

Section: Introductionmentioning

confidence: 99%

Humanization of Yeasts for Glycan-Type End-Products

Li¹,

Shen²,

Chen³

et al. 2022

Front. Microbiol.

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Yeasts are often considered microorganisms for producing human therapeutic glycosylated end-products at an industrial scale. However, the products with non-humanized glycans limited their usage. Therefore, various methods to develop humanized glycosylated end-products have been widely reported in yeasts. To make full use of these methods, it is necessary to summarize the present research to find effective approaches to producing humanized products. The present research focuses on yeast species selection, glycosyltransferase deletion, expression of endoglycosidase, and expression of proteins with galactosylated and or sialylated glycans. Nevertheless, the yeasts will have growth defects with low bioactivity when the key enzymes are deleted. It is necessary to express the corresponding repairing protein. Compared with N-glycosylation, the function of yeast protein O-glycosylation is not well-understood. Yeast proteins have a wide variety of O-glycans in different species, and it is difficult to predict glycosylation sites, which limits the humanization of O-glycosylated yeast proteins. The future challenges include the following points: there are still many important potential yeasts that have never been tried to produce glycosylated therapeutic products. Their glycosylation pathway and related mechanisms for producing humanized glycosylated proteins have rarely been reported. On the other hand, the amounts of key enzymes on glycan pathways in human beings are significantly more than those in yeasts. Therefore, there is still a challenge to produce a large body of humanized therapeutic end-products in suitable yeast species, especially the protein with complex glycans. CRISPR-Cas9 system may provide a potential approach to address the important issue.

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Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2009–2010

Harvey

2014

Mass Spectrometry Reviews

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This review is the sixth update of the original article published in 1999 on the application of MALDI mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings coverage of the literature to the end of 2010. General aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, arrays and fragmentation are covered in the first part of the review and applications to various structural typed constitutes the remainder. The main groups of compound that are discussed in this section are oligo and polysaccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals. Many of these applications are presented in tabular form. Also discussed are medical and industrial applications of the technique, studies of enzyme reactions and applications to chemical synthesis.

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Structure of glycosylated Cu/Zn-superoxide dismutase from Kluyveromyces yeast NBIMCC 1984

Cited by 5 publications

References 38 publications

N-Glycosylation profiling of recombinant mouse extracellular superoxide dismutase produced in Chinese hamster ovary cells

N-Glycosylation profiling of recombinant mouse extracellular superoxide dismutase produced in Chinese hamster ovary cells

Humanization of Yeasts for Glycan-Type End-Products

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2009–2010

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