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Current Opinion in Biotechnology Oct 2021Protein glycosylation is the most diverse and omnipresent protein modification. Glycosylation provides glycoproteins with important structural and functional properties... (Review)
Review
Protein glycosylation is the most diverse and omnipresent protein modification. Glycosylation provides glycoproteins with important structural and functional properties to facilitate critical biological processes. Despite the significance of protein glycosylation, the investigation of glycoproteome, especially O-linked glycoproteome, remains elusive due to the lack of a comprehensive methodology to conform with the diversity of O-linked glycoforms of O-linked glycoproteins. In recent years, mass spectrometry has become an indispensable tool for the characterization of O-linked glycosylated proteins across biological systems. We herein highlight the recent developments in MS-based O-linked glycoproteomic technologies, quantitative data acquisition strategy and bioinformatic tools, with a special focus on mucin-type O-linked glycosylation.
Topics: Glycoproteins; Glycosylation; Protein Processing, Post-Translational; Proteome
PubMed: 34358979
DOI: 10.1016/j.copbio.2021.07.006 -
Plant Science : An International... Oct 2015Asparagine (N)-linked protein glycosylation is one of the most crucial, prevalent, and complex co- and post-translational protein modifications. It plays a pivotal role... (Review)
Review
Asparagine (N)-linked protein glycosylation is one of the most crucial, prevalent, and complex co- and post-translational protein modifications. It plays a pivotal role in protein folding, quality control, and endoplasmic reticulum (ER)-associated degradation (ERAD) as well as in protein sorting, protein function, and in signal transduction. Furthermore, glycosylation modulates many important biological processes including growth, development, morphogenesis, and stress signaling processes. As a consequence, aberrant or altered N-glycosylation is often associated with reduced fitness, diseases, and disorders. The initial steps of N-glycan synthesis at the cytosolic side of the ER membrane and in the lumen of the ER are highly conserved. In contrast, the final N-glycan processing in the Golgi apparatus is organism-specific giving rise to a wide variety of carbohydrate structures. Despite our vast knowledge on N-glycans in yeast and mammals, the modus operandi of N-glycan signaling in plants is still largely unknown. This review will elaborate on the N-glycosylation biosynthesis pathway in plants but will also critically assess how N-glycans are involved in different signaling cascades, either active during normal development or upon abiotic and biotic stresses.
Topics: Glycosylation; Plant Development; Plants; Polysaccharides; Protein Processing, Post-Translational; Signal Transduction; Stress, Physiological
PubMed: 26398792
DOI: 10.1016/j.plantsci.2015.06.023 -
Nature Methods Jan 2019
Topics: Animals; Glycosylation; Nucleic Acid Hybridization; Zebrafish
PubMed: 30573827
DOI: 10.1038/s41592-018-0284-8 -
Advances in Carbohydrate Chemistry and... 2022In the post-genomic era, post- and co-translational modifications (P/C-TM) of proteins are known as the more essential elements for the activation of protein function.... (Review)
Review
In the post-genomic era, post- and co-translational modifications (P/C-TM) of proteins are known as the more essential elements for the activation of protein function. Among these protein modifications, glycosylation is one of the most abundant modifications in eukaryotic cells. The synthesis of glycoproteins with uniform glycan structures is essential for functional analysis of glycoproteins and biochemical research. For that purpose, chemical methods to synthesize glycoproteins with chemically uniform glycan structures have been developed. In this review, we highlight our recent advances in the preparation of homogeneous glycoproteins. Especially, we introduce both semi-synthesis and chemical synthesis of glycoproteins along with semi-synthesis of various complex-type N-glycans for the solid-phase synthesis of glycopeptide-thioesters.
Topics: Glycoproteins; Polysaccharides; Glycosylation; Glycopeptides; Protein Processing, Post-Translational
PubMed: 36435534
DOI: 10.1016/bs.accb.2022.09.004 -
Current Opinion in Biotechnology Dec 2022The commercial value of specialty carbohydrates and glycosylated compounds has sparked considerable interest in the synthetic potential of carbohydrate-active enzymes... (Review)
Review
The commercial value of specialty carbohydrates and glycosylated compounds has sparked considerable interest in the synthetic potential of carbohydrate-active enzymes (CAZymes). Protein engineering methods have proven to be highly successful in expanding the range of glycosylation reactions that these enzymes can perform efficiently and cost-effectively. The past few years have witnessed meaningful progress in this area, largely due to a sharper focus on the understanding of structure-function relationships and mechanistic intricacies. Here, we summarize recent studies that demonstrate how protein engineers have become much better at traversing the fitness landscape of CAZymes through mutational bridges that connect the different activity types.
Topics: Carbohydrates; Glycosylation; Proteins; Enzymes
PubMed: 36156353
DOI: 10.1016/j.copbio.2022.102804 -
Journal of Proteome Research Jun 2023Glycosylation is one of the most common and important protein modifications, and it regulates the properties and functions of a wide range of proteins. Aberrant... (Review)
Review
Glycosylation is one of the most common and important protein modifications, and it regulates the properties and functions of a wide range of proteins. Aberrant glycosylation is directly related to human diseases. Recently, with the advancement of mass spectrometry (MS) instrumentation and MS-based glycoproteomic methods, global characterization of glycoproteins in complex biological samples has become possible. Using quantitative proteomics, the abundance of glycoproteins in different samples can be quantified, which provides a wealth of information to further our understanding of protein functions, cellular activities, and the molecular mechanisms of diseases. In this review, we discuss quantitative proteomic methods used for comprehensive analysis of protein glycosylation, and cover the applications of quantitative glycoproteomics to unveil the properties and functions of glycoproteins and their association with various diseases. It is expected that quantitative proteomic methods will be extensively applied to explore the role of protein glycosylation in complex biological systems, and to identify glycoproteins as biomarkers for disease detection and as therapeutic targets for disease treatment.
Topics: Humans; Proteomics; Glycoproteins; Glycosylation; Protein Processing, Post-Translational; Mass Spectrometry
PubMed: 37010087
DOI: 10.1021/acs.jproteome.3c00015 -
Mass Spectrometry Reviews Aug 2019Protein glycosylation is ubiquitous in biological systems and plays essential roles in many cellular events. Global and site-specific analysis of glycoproteins in... (Review)
Review
Protein glycosylation is ubiquitous in biological systems and plays essential roles in many cellular events. Global and site-specific analysis of glycoproteins in complex biological samples can advance our understanding of glycoprotein functions and cellular activities. However, it is extraordinarily challenging because of the low abundance of many glycoproteins and the heterogeneity of glycan structures. The emergence of mass spectrometry (MS)-based proteomics has provided us an excellent opportunity to comprehensively study proteins and their modifications, including glycosylation. In this review, we first summarize major methods for glycopeptide/glycoprotein enrichment, followed by the chemical and enzymatic methods to generate a mass tag for glycosylation site identification. We next discuss the systematic and quantitative analysis of glycoprotein dynamics. Reversible protein glycosylation is dynamic, and systematic study of glycoprotein dynamics helps us gain insight into glycoprotein functions. The last part of this review focuses on the applications of MS-based proteomics to study glycoproteins in different biological systems, including yeasts, plants, mice, human cells, and clinical samples. Intact glycopeptide analysis is also included in this section. Because of the importance of glycoproteins in complex biological systems, the field of glycoproteomics will continue to grow in the next decade. Innovative and effective MS-based methods will exponentially advance glycoscience, and enable us to identify glycoproteins as effective biomarkers for disease detection and drug targets for disease treatment. © 2019 Wiley Periodicals, Inc. Mass Spec Rev 9999: XX-XX, 2019.
Topics: Animals; Glycomics; Glycopeptides; Glycoproteins; Glycosylation; Humans; Mass Spectrometry; Polysaccharides; Proteomics
PubMed: 30605224
DOI: 10.1002/mas.21586 -
Frontiers in Cellular and Infection... 2020In recent years, protein glycosylation in pathogenic bacteria has attracted more and more attention, and accumulating evidence indicated that this type of... (Review)
Review
In recent years, protein glycosylation in pathogenic bacteria has attracted more and more attention, and accumulating evidence indicated that this type of posttranslational modification is involved in many physiological processes. The NleB from several enteropathogenic bacteria species as well as SseK from are type III secretion system effectors, which have an atypical N-acetylglucosamine (N-GlcNAc) transferase activity that specifically modified a conserved arginine in TRADD, FADD, and RIPK1. NleB/SseKs GlcNAcylation of death domain proteins abrogates homotypic and heterotypic death receptors/adaptors interactions, thereby blocking an important antimicrobial host response. Interestingly, NleB/SseKs could also GlcNAcylate themselves, and self-GlcNAcylation of NleB, SseK1, and SseK3 are crucial for their biological activity during infection. In addition, EarP (EF-P specific arginine rhamnosyl transferase for Posttranslational activation) catalyzes arginine rhamnosylation of translation elongation factor P (EF-P). Importantly, this kind of N-linked protein glycosylation is not only important for EF-P dependent rescue of polyproline stalled ribosomes but also for pathogenicity in and other clinically relevant bacteria. Glycosylation of arginine is unique because the guanidine group of arginine has a high acid dissociation constant value and representing an extremely poor nucleophile. Recently, the crystal structures of NleB, SseKs, EarP, arginine GlcNAcylated death domain-containing proteins, NleB/FADD-DD, and EarP/EF-P/dTDP-β-L-rhamnose were solved by our group and other groups, revealing the unique catalytic mechanisms. In this review, we provide detailed information about the currently known arginine glycosyltransferases and their potential catalytic mechanisms.
Topics: Arginine; Bacterial Proteins; Catalysis; Fas-Associated Death Domain Protein; Glycosylation; Pseudomonas aeruginosa; Receptor-Interacting Protein Serine-Threonine Kinases; TNF Receptor-Associated Death Domain Protein
PubMed: 32411621
DOI: 10.3389/fcimb.2020.00185 -
Clinical & Translational Oncology :... Oct 2022Endometrial cancer (EC) is one of the most common tumors in the female reproductive system, which seriously threatens women's health, particularly in developed... (Review)
Review
Endometrial cancer (EC) is one of the most common tumors in the female reproductive system, which seriously threatens women's health, particularly in developed countries. 13% of the patients with EC have a poor prognosis due to recurrence and metastasis. Therefore, identifying good predictive biomarkers and therapeutic targets is critical to enable the early detection of metastasis and improve the prognosis. For decades, extensive studies had focused on glycans and glycoproteins in the progression of cancer. The types of glycans that are covalently attached to the polypeptide backbone, usually via nitrogen or oxygen linkages, are known as N‑glycans or O‑glycans, respectively. The degree of protein glycosylation and the aberrant changes in the carbohydrate structures have been implicated in the extent of tumorigenesis and reported to play a critical role in regulating tumor invasion, metabolism, and immunity. This review summarizes the essential biological role of glycosylation in EC, with a focus on the recent advances in glycomics and glycosylation markers, highlighting their implications in the diagnosis and treatment of EC.
Topics: Biomarkers; Endometrial Neoplasms; Female; Glycomics; Glycoproteins; Glycosylation; Humans; Polysaccharides
PubMed: 35752750
DOI: 10.1007/s12094-022-02858-z -
Clinical Chemistry and Laboratory... Mar 2019Glycosylation is among the most important post-translational modifications for proteins and is of intrinsic complex character compared with DNAs and naked proteins.... (Review)
Review
Glycosylation is among the most important post-translational modifications for proteins and is of intrinsic complex character compared with DNAs and naked proteins. Indeed, over 50%-70% of proteins in circulation are glycosylated, and the "sweet attachments" have versatile structural and functional implications. Both the configuration and composition of the attached glycans affect the biological activities of consensus proteins significantly. Glycosylation is generated by complex biosynthetic pathways comprising hundreds of glycosyltransferases, glycosidases, transcriptional factors, transporters and the protein backbone. In addition, lack of direct genetic templates and glyco-specific antibodies such as those commonly used in DNA amplification and protein capture makes research on glycans and glycoproteins even more difficult, thus resulting in sparse knowledge on the pathophysiological implications of glycosylation. Fortunately, cutting-edge technologies have afforded new opportunities and approaches for investigating cancer-related glycosylation. Thus, glycans as well as aberrantly glycosylated protein-based cancer biomarkers have been increasingly recognized. This mini-review highlights the most recent developments in glyco-biomarker studies in an effort to discover clinically relevant cancer biomarkers using advanced analytical methodologies such as mass spectrometry, high-performance liquid chromatographic/ultra-performance liquid chromatography, capillary electrophoresis, and lectin-based technologies. Recent clinical-centered glycobiological studies focused on determining the regulatory mechanisms and the relation with diagnostics, prognostics and even therapeutics are also summarized. These studies indicate that glycomics is a treasure waiting to be mined where the growth of cancer-related glycomics and glycoproteomics is the next great challenge after genomics and proteomics.
Topics: Biomarkers, Tumor; Glycosylation; Humans; Neoplasms
PubMed: 30138110
DOI: 10.1515/cclm-2018-0379