• Journal of Microbiology and Biotechnology
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• v.30 no.7
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• pp.1092-1096
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• 2020

YjiC, a glycosyltransferase from Bacillus licheniformis, is a well-known versatile enzyme for glycosylation of diverse substrates. Although a number of O-glycosylated products have been produced using YjiC, no report has been updated for nucleophilic N-, S-, and C- glycosylation. Here, we report the additional functional capacity of YjiC for nucleophilic N- and S- glycosylation using a broad substrate spectrum including UDP-α-D-glucose, UDP-N-acetyl glucosamine, UDP-N-acetylgalactosamine, UDP-α-D-glucuronic acid, TDP-α-L-rhamnose, TDP-α-D-viosamine, and GDP-α-L-fucose as donor and various amine and thiol groups containing natural products as acceptor substrates. The results revealed YjiC as a promiscuous enzyme for conjugating diverse sugars at amine and thiol functional groups of small molecules applicable for generating glycofunctionalized chemical diversity libraries. The glycosylated products were analyzed using HPLC and LC/MS and compared with previous reports.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.721-724
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• 2000

The possibility of the enzymatic in vitro glycosylation using peptide-N-glycosidase F was examined. Oligosaccharide chains in the glycoproteins are important for the biological activity, solubility, immunogenecity, recognition, and prevention of degradation. After 4 h incubation of deglycosylated glycoprotein with excess glucose oligomer and ammonia in acetone at $50^{\circ}C$, upper shift of protein band was observed on SDS-PAGE. And the different deglycosylation characteristics of glucose oxidase and fetuin were investigated.

• BMB Reports
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• v.45 no.5
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• pp.317-322
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• 2012

2-deoxy-D-glucose(2DG)-caused endoplasmic reticulum (ER) stress inhibits protein phosphorylation at tyrosine residues. However, the accurate regulatory mechanisms, which determine the inflammatory response of chondrocytes to ER stress via protein tyrosine phosphorylation, have not been systematically evaluated. Thus, in this study, we examined whether protein phosphorylation at tyrosine residues can modulate the expression and glycosylation of COX-2, which is reduced by 2DG-induced ER stress. We observed that protein tyrosine phosphatase (PTP) inhibitors, sodium orthovanadate (SOV), and phenylarsine oxide (PAO) significantly decreased expression of ER stress inducible proteins, glucose-regulated protein 94 (GRP94), and CCAAT/ enhancer-binding-protein- related gene (GADD153), which was induced by 2DG. In addition, we demonstrated that SOV and PAO noticeably restored the expression and glycosylation of COX-2 after treatment with 2DG. These results suggest that protein phosphorylation of tyrosine residues plays an important role in the regulation of expression and glycosylation during 2DG-induced ER stress in rabbit articular chondrocytes.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.24 no.1
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• pp.10-16
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• 2024

Congenital Disorders of Glycosylation (CDG) represent a complex group of inherited metabolic disorders resulting from defects in multiple pathways of glycosylation, a critical biochemical process for protein functionality and cellular communication. This review provides a comprehensive overview of CDG, including its history, epidemiology, classification, diagnostic complexities, and therapeutic developments. Despite advancements in understanding CDG and identifying over 160 subtypes, challenges remain due to the diverse clinical manifestations and multi-systemic involvement. Targeted therapy is available for only a few CDGs, but promising treatments are being investigated. Ongoing research is vital to developing targeted treatments and improving patient outcomes.

• Proceedings of the Korean Biophysical Society Conference
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• 1997.07a
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• pp.25-25
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• 1997

Cyclic nucleotide-gated channels (CNGC's) contain a putative N-glycosylation site (Asn-X-Ser/Thr) in the linker regions connecting the fourth transmembrane domain (S4) and the ion conduction pore (P-region). This putative N-glycosylation site is highly conserved and thus found in many different CNGC in various organisms, from fruit to human.(omitted)

• BMB Reports
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• v.54 no.11
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• pp.541-544
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• 2021

Protein glycosylation is a common post-translational modification found in all living organisms. This modification in bacterial pathogens plays a pivotal role in their infectious processes including pathogenicity, immune evasion, and host-pathogen interactions. Importantly, many key proteins of host immune systems are also glycosylated and bacterial pathogens can notably modulate glycosylation of these host proteins to facilitate pathogenesis through the induction of abnormal host protein activity and abundance. In recent years, interest in studying the regulation of host protein glycosylation caused by bacterial pathogens is increasing to fully understand bacterial pathogenesis. In this review, we focus on how bacterial pathogens regulate remodeling of host glycoproteins during infections to promote the pathogenesis.

• Mass Spectrometry Letters
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• v.15 no.3
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• pp.121-140
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• 2024

Proteoform diversity is greatly increased by glycosylation, the primary post-translational modification of proteins. Glycans, also known as oligosaccharides, are molecules that are essential to almost all living things. They can affect protein folding and functionality, modulate cell-cell interactions, and support the proliferation of numerous diseases when they are found on cell surfaces or bound to proteins. A thorough understanding of their fundamental structure is necessary to gain insight into their characteristics and functions. But a major obstacle is the structural intricacy of glycans by design. The stereochemistry and regiochemistry of carbohydrates vary and are frequently branched. Because of its superior sensitivity and the abundance of fragmentation information it can provide, mass spectrometry is now the method of choice for glycan and glycopeptide analysis. Differentiating between the structures of isomeric and isobaric glycopeptides, however, presents a difficulty for MS-based characterization. Ion mobility plus mass spectrometry (IM-MS) has become a very promising new method for glycan research in recent years. Recent developments in the growing discipline of glycosylation analysis utilizing IM-MS are outlined in this review, with a focus on the MS methodology and its ability to resolve isomeric glycans.

• IMMUNE NETWORK
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• v.9 no.2
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• pp.58-63
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• 2009

Background: T cell immunoglobulin and mucin domain containing 3 protein (Tim-3) expressed on terminally differentiated Th1 cells plays a suppressive role in Th1-mediated immune responses. Recently, it has been shown that N-glycosylation affects the binding activity of the Tim-3-Ig fusion protein to its ligand, galectin-9, but the binding properties of non-glycosylated Tim-3 on $CD4^+CD25^+$T cells has not been fully examined. In this study, we produced recombinant Tim-3-Ig fusion proteins in different cellular sources and its N-glycosylation mutant forms to evaluate their binding activities to $CD4^+CD25^+$T cells. Methods: We isolated and cloned Tim-3 cDNA from BALB/C mouse splenocytes. Then, we constructed a mammalian expression vector and a prokaryotic expression vector for the Tim-3-Ig fusion protein. Using a site directed mutagenesis method, plasmid vectors for Tim-3-Ig N-glycosylation mutant expression were produced. The recombinant protein was purified by protein A sepharose column chromatography. The binding activity of Tim-3-Ig fusion protein to $CD4^+CD25^+$T cells was analyzed using flow cytometry. Results: We found that the nonglycosylated Tim-3-Ig fusion proteins expressed in bacteria bound to $CD4^+CD25^+$T cells similarly to the glycosylated Tim-3-Ig protein produced in CHO cells. Further, three N-glycosylation mutant forms (N53Q, N100Q, N53/100Q) of Tim-3-Ig showed similar binding activities to those of wild type glycosylated Tim-3-Ig. Conclusion: Our results suggest that N-glycosylation of Tim-3 may not affect its binding activity to ligands expressed on $CD4^+CD25^+$T cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.29-32
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• 2004

Inhibition of the early N-glycosylation process in the endoplasmic reticulum prevents the activation of tyrosinase, a key enzyme for melanin biosynthesis. This work aims at evaluating the increased activity of N-glycosylation inhibitors in vitro b, employing a nano-sized pH-sensitive liposome as a delivery carrier. Melexsome, a pH-sensitive nano carrier loaded with glycosylation inhibitos, was prepared by the hydration method with phospholipids and cholresterol-based amphiphiles. Inhibitory effects of Melexsome on the N-glycosylation process were evaluated by EndoH ＆ PNGaseF digestion and the western blotting. Melanin synthesis was also monitored after treatment with Melexsome Interestingly, Melexsome effectively increased the efficacy of N-glycosylation inhibitors. Melexsome was also much more efficiently translocated into the cytoplasm as observed in CLSM. These results demonstrated that the amphiphilic lipid-based pH-sensitive nano-carriers could be, used as an efficient delivery system for N-glycosylation inhibitor to enhance the effects of skin whitening cosmetics.

• Bulletin of the Korean Chemical Society
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• v.25 no.1
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• pp.139-142
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• 2004