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Carbohydrate Research Apr 2022Monoclonal antibodies (mAbs) are amazingly successful in treating diseases such as various types of cancers, immune disorders, and infections. Currently, mAbs are... (Review)
Review
Monoclonal antibodies (mAbs) are amazingly successful in treating diseases such as various types of cancers, immune disorders, and infections. Currently, mAbs are considered the most versatile therapeutic available. Seminal scientific progress over the last decade on elucidating the functional role of antibody N-glycosylation revealed the significance of this post-translational modification on the stability, half-life, and biological activity of antibodies. Therefore, antibody N-glycosylation became a key aspect of biopharmaceutical mAb production in large-scale manufacturing processes. This Mini-Review summarizes the recent scientific attempts of producing mAbs in glycoengineered animal-, plant-, and yeast cell lines. Furthermore, we also describe novel approaches for analyzing mAb N-glycosylation.
Topics: Animals; Antibodies, Monoclonal; Biotechnology; Glycosylation; Neoplasms; Protein Processing, Post-Translational
PubMed: 35344785
DOI: 10.1016/j.carres.2022.108541 -
Current Opinion in Structural Biology Jun 2019Bacterial protein glycosylation (BPG) comes in all variations of form and manifestation: N-versus O-linked, dedicated versus broad-spectrum, sequential versus en bloc... (Review)
Review
Bacterial protein glycosylation (BPG) comes in all variations of form and manifestation: N-versus O-linked, dedicated versus broad-spectrum, sequential versus en bloc glycan addition, cytoplasmic versus extracytoplasmic. Here, I evaluate and address recent advances in the field of O-linked BPG focusing on selected systems of notoriety and in which significant advances have occurred of late.
Topics: Bacteria; Bacterial Proteins; Flagellin; Glycosylation; Hexosyltransferases; Membrane Proteins; Oxygen
PubMed: 31078896
DOI: 10.1016/j.sbi.2019.03.020 -
Molecular Biology Reports Aug 2022N-linked protein glycosylation is an essential co-and posttranslational protein modification that occurs in all three domains of life; the assembly of N-glycans follows... (Review)
Review
N-linked protein glycosylation is an essential co-and posttranslational protein modification that occurs in all three domains of life; the assembly of N-glycans follows a complex sequence of events spanning the (Endoplasmic Reticulum) ER and the Golgi apparatus. It has a significant impact on both physicochemical properties and biological functions. It plays a significant role in protein folding and quality control, glycoprotein interaction, signal transduction, viral attachment, and immune response to infection. Glycoengineering of protein employed for improving protein properties and plays a vital role in the production of recombinant glycoproteins and struggles to humanize recombinant therapeutic proteins. It considers an alternative platform for biopharmaceuticals production. Many immune proteins and antibodies are glycosylated. Pathogen's glycoproteins play vital roles during the infection cycle and their expression of specific oligosaccharides via the N-glycosylation pathway to evade detection by the host immune system. This review focuses on the aspects of N-glycosylation processing, glycoengineering approaches, their role in viral attachment, and immune responses to infection.
Topics: Endoplasmic Reticulum; Glycoproteins; Glycosylation; Golgi Apparatus; Humans; Polysaccharides; Recombinant Proteins; Virus Diseases
PubMed: 35364718
DOI: 10.1007/s11033-022-07359-4 -
Expert Review of Proteomics 2023Hepatocellular carcinoma (HCC) represents a significant burden globally, which ranks sixth among the most frequently diagnosed cancers and stands as the third leading... (Review)
Review
INTRODUCTION
Hepatocellular carcinoma (HCC) represents a significant burden globally, which ranks sixth among the most frequently diagnosed cancers and stands as the third leading cause of cancer-related mortality. Glycoproteomics, as an important branch of proteomics, has already made significant achievements in the field of HCC research. Aberrant protein glycosylation has shown to promote the malignant transformation of hepatocytes by modulating a wide range of tumor-promoting signaling pathways. The glycoproteome provides valuable information for understanding cancer progression, tumor immunity, and clinical outcome, which could serve as potential diagnostic, prognostic, and therapeutic tools in HCC.
AREAS COVERED
In this review, recent advances of glycoproteomics contribute to clinical applications (diagnosis and prognosis) and molecular mechanisms (hepatocarcinogenesis, progression, stemness and recurrence, and drug resistance) of HCC are summarized.
EXPERT OPINION
Glycoproteomics shows promise in HCC, enhancing early detection, risk stratification, and personalized treatments. Challenges include sample heterogeneity, diverse glycans structures, sensitivity issues, complex workflows, limited databases, and incomplete understanding of immune cell glycosylation. Addressing these limitations requires collaborative efforts, technological advancements, standardization, and validation studies. Future research should focus on targeting abnormal protein glycosylation therapeutically. Advancements in glycobiomarkers and glycosylation-targeted therapies will greatly impact HCC diagnosis, prognosis, and treatment.
Topics: Humans; Carcinoma, Hepatocellular; Liver Neoplasms; Glycoproteins; Glycosylation; Polysaccharides
PubMed: 37882248
DOI: 10.1080/14789450.2023.2265064 -
Trends in Molecular Medicine Feb 2024Protein glycosylation controls cell-cell and cell-extracellular matrix (ECM) communication in immune, vascular, and inflammatory processes, underlining the critical role... (Review)
Review
Protein glycosylation controls cell-cell and cell-extracellular matrix (ECM) communication in immune, vascular, and inflammatory processes, underlining the critical role of this process in the identification of disease biomarkers and the design of novel therapies. Emerging evidence highlights the critical role of blood cell glycosylation in the pathophysiology of atherosclerosis (ATH) and myocardial infarction (MI). Here, we review the role of glycosylation in the interplay between blood cells, particularly erythrocytes, and endothelial cells (ECs), highlighting the involvement of this critical post/cotranslational modification in settings of cardiovascular disease (CVD). Importantly, we focus on emerging preclinical studies and clinical trials based on glycan-targeted drugs to validate their therapeutic potential. These findings may help establish new trends in preventive medicine and delineate novel targeted therapies in CVD.
Topics: Humans; Glycosylation; Endothelial Cells; Myocardial Infarction; Atherosclerosis; Blood Cells
PubMed: 38142190
DOI: 10.1016/j.molmed.2023.11.013 -
Methods in Molecular Biology (Clifton,... 2023While the knowledge of protein structure and function has seen vast advances in previous decades, the understanding of how their posttranslational modifications, such as...
While the knowledge of protein structure and function has seen vast advances in previous decades, the understanding of how their posttranslational modifications, such as glycosylations, influence their structure and function remains poor. However, advances in in silico methodologies to study glycosylations in recent past have enabled us to study this and understand the role of glycosylations in protein structure and function in ways that would not be possible by conventional experimental methods. In this chapter, we will demonstrate how to leverage these methodologies to study glycoproteins and their structural and dynamic properties using molecular modelling techniques.
Topics: Glycoproteins; Glycosylation; Models, Molecular; Protein Processing, Post-Translational
PubMed: 36662478
DOI: 10.1007/978-1-0716-2946-8_21 -
Current Opinion in Biotechnology Dec 2019For a long time, glycoprotein production has been limited by the inherent properties of production hosts. Glycosylation of biopharmaceuticals has been regarded as a... (Review)
Review
For a long time, glycoprotein production has been limited by the inherent properties of production hosts. Glycosylation of biopharmaceuticals has been regarded as a necessary evil, often needed for protein folding or function, but also a source of heterogeneity, complicating downstream processing and product characterization. This has strongly determined the choice of production hosts. Over the last few decades, numerous glycoengineering efforts have helped solving this problem. Moreover, insights from fundamental studies have made it possible to improve therapeutic protein functionality through careful glycoengineering. Here, we will focus on how production host and in vitro glycoengineering approaches allow to design biopharmaceuticals with glycans that impart improved functionality. An important branch of research explores how glycosylation can be tuned to improve pharmacokinetics and reduce glycan heterogeneity of therapeutics. Furthermore, antibody glycoengineering to obtain homogeneous, defined glycan structures has been a major focus. An example of this is the production of Fc glycans without core fucose, exhibiting tremendously improved Antibody-Dependent Cell Cytotoxicity (ADCC). In the last part, glycoforms that allow for improved (subcellular) targeting and cellular uptake, a field that opens possibilities for enzyme replacement therapies and vaccine development, will be highlighted.
Topics: Antibodies; Biological Products; Glycoproteins; Glycosylation; Polysaccharides
PubMed: 30554064
DOI: 10.1016/j.copbio.2018.11.017 -
Cellular and Molecular Life Sciences :... Jul 2015Asparagine (N)-linked protein glycosylation, which takes place in the eukaryotic endoplasmic reticulum (ER), is important for protein folding, quality control and the... (Review)
Review
Asparagine (N)-linked protein glycosylation, which takes place in the eukaryotic endoplasmic reticulum (ER), is important for protein folding, quality control and the intracellular trafficking of secretory and membrane proteins. It is known that, during N-glycosylation, considerable amounts of lipid-linked oligosaccharides (LLOs), the glycan donor substrates for N-glycosylation, are hydrolyzed to form free N-glycans (FNGs) by unidentified mechanisms. FNGs are also generated in the cytosol by the enzymatic deglycosylation of misfolded glycoproteins during ER-associated degradation. FNGs derived from LLOs and misfolded glycoproteins are eventually merged into one pool in the cytosol and the various glycan structures are processed to a near homogenous glycoform. This article summarizes the current state of our knowledge concerning the formation and catabolism of FNGs.
Topics: Asparagine; Biosynthetic Pathways; Endoplasmic Reticulum; Glycosylation; Hydrolysis; Lipopolysaccharides; Models, Biological; Polysaccharides; Protein Folding
PubMed: 25772500
DOI: 10.1007/s00018-015-1881-7 -
Mass Spectrometry-Based Chemical and Enzymatic Methods for Global Analysis of Protein Glycosylation.Accounts of Chemical Research Aug 2018Glycosylation is one of the most common protein modifications, and it is essential for mammalian cell survival. It often determines protein folding and trafficking, and... (Review)
Review
Glycosylation is one of the most common protein modifications, and it is essential for mammalian cell survival. It often determines protein folding and trafficking, and regulates nearly every extracellular activity, including cell-cell communication and cell-matrix interactions. Aberrant protein glycosylation events are hallmarks of human diseases such as cancer and infectious diseases. Therefore, glycoproteins can serve as effective biomarkers for disease detection and targets for drug and vaccine development. Despite the importance of glycoproteins, global analysis of protein glycosylation (either glycoproteins or glycans) in complex biological samples has been a daunting task, and here we mainly focus on glycoprotein analysis using mass spectrometry (MS)-based bottom-up proteomics. Although the emergence of MS-based proteomics has provided a great opportunity to analyze glycoproteins globally, the low abundance of many glycoproteins and the heterogeneity of glycans dramatically increase the technical difficulties. In order to overcome these obstacles, considerable progress has been made in recent years, which has contributed to comprehensive analysis of glycoproteins. In our lab, we developed effective MS-based chemical and enzymatic methods to (1) globally analyze glycoproteins in complex biological samples, (2) target glycoproteins specifically on the surface of human cells, (3) systematically quantify glycoprotein and surface glycoprotein dynamics (the abundance changes of glycoproteins as a function of time), and (4) selectively characterize glycoproteins with a particular and important glycan. In this Account, we first briefly describe the glycopeptide/protein enrichment methods in the literature and then discuss the developments of boronic acid-based methods to enrich glycopeptides for large-scale analysis of protein glycosylation. Boronic acids can form reversible covalent interactions with sugars, but the low binding affinity of normal boronic acid-based methods prevents us from capturing glycoproteins with low abundance, which often contain more valuable information. We enhanced the boronic acid-glycan interactions by using a boronic acid derivative (benzoboroxole) and conjugating it onto a dendrimer to allow synergistic interactions between the boronic acid derivative and sugars. The new method is capable of globally analyzing protein glycosylation with site and glycan structure information, especially for those with low abundance. In the next part, we discuss the combination of metabolic labeling, click chemistry and enzymatic reactions, and MS-based proteomics as a very powerful approach for surface glycoproteome analysis in human cells. The methods enable us to specifically identify surface glycoproteins and to quantify their abundance changes and dynamics together with quantitative proteomics. The last section of this Account focuses on chemical and enzymatic methods to study glycoproteins containing a particular and important glycan (the Tn antigen, i.e., O-GalNAc). Although not comprehensive, this Account provides an overview of chemical and enzymatic methods to characterize protein glycosylation in combination with MS-based proteomics. These methods will have extensive applications in the fields of biology and biomedicine, which will lead to a better understanding of glycoprotein functions and the molecular mechanisms of diseases. Eventually, glycoproteins will be identified as effective biomarkers for disease detection and drug targets for disease treatment.
Topics: Antigens, Tumor-Associated, Carbohydrate; Boronic Acids; Click Chemistry; Glycosylation; Humans; Mass Spectrometry; Membrane Glycoproteins; Polysaccharides; Protein Processing, Post-Translational; Proteome; Proteomics
PubMed: 30011186
DOI: 10.1021/acs.accounts.8b00200 -
Pflugers Archiv : European Journal of... Jul 2021Proper protein glycosylation is critical to normal cardiomyocyte physiology. Aberrant glycosylation can alter protein localization, structure, drug interactions, and... (Review)
Review
Proper protein glycosylation is critical to normal cardiomyocyte physiology. Aberrant glycosylation can alter protein localization, structure, drug interactions, and cellular function. The in vitro differentiation of human pluripotent stem cells into cardiomyocytes (hPSC-CM) has become increasingly important to the study of protein function and to the fields of cardiac disease modeling, drug testing, drug discovery, and regenerative medicine. Here, we offer our perspective on the importance of protein glycosylation in hPSC-CM. Protein glycosylation is dynamic in hPSC-CM, but the timing and extent of glycosylation are still poorly defined. We provide new data highlighting how observed changes in hPSC-CM glycosylation may be caused by underlying differences in the protein or transcript abundance of enzymes involved in building and trimming the glycan structures or glycoprotein gene products. We also provide evidence that alternative splicing results in altered sites of glycosylation within the protein sequence. Our findings suggest the need to precisely define protein glycosylation events that may have a critical impact on the function and maturation state of hPSC-CM. Finally, we provide an overview of analytical strategies available for studying protein glycosylation and identify opportunities for the development of new bioinformatic approaches to integrate diverse protein glycosylation data types. We predict that these tools will promote the accurate assessment of protein glycosylation in future studies of hPSC-CM that will ultimately be of significant experimental and clinical benefit.
Topics: Animals; Glycosylation; Humans; Myocytes, Cardiac; Pluripotent Stem Cells; Proteins
PubMed: 33830329
DOI: 10.1007/s00424-021-02554-x