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New Insights into Protein Glycosylation

A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Chemical Biology".

Deadline for manuscript submissions: closed (10 May 2022) | Viewed by 52341

Special Issue Editor


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Guest Editor
School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka 422-8526, Japan
Interests: glycoproteomics; protein quality control; protein folding; EGF repeats; O-glycosylation; O-glucosylation; mass spectrometry; glycosyltransferase; notch signaling; biohchemistry; glycobiology; cancer biology
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

To understand life in an integrated manner, it is essential to understand protein glycosylation: a growing and high-impact field exploring the structure and function of the carbohydrates that clothe most living cells. In this Special Issue, we hope to take an interdisciplinary approach, particularly highlighting work in chemistry and biology, to summarize the current status and discuss the future development and prospects of protein glycosylation in terms of its functions and analytical methods. As described on the website of Molecules, we will accept regular original Articles and Reviews. (Please note that the former includes Short communications of preliminary, but significant, results.) Protein glycosylation in extracellular or membrane proteins includes N-linked glycosylation and mucin-type O-linked glycosylation starting with GalNAc in addition to O-Fuc, O-Gal, O-Glc, O-GlcNAc, O-Man and O-Xyl, and C-Man. There are also a variety of nucleocytoplasmic proteins modified with O-GlcNAc by OGT. Their functions are involved in diverse biological processes such as cellular signaling, immunity, neural function, pathological conditions such as cancer, and viral infections, including COVID-19.

Prof. Dr. Hideyuki Takeuchi
Guest Editor

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Keywords

  • O-glycosylation
  • N-glycosylation
  • C-mannosylation
  • glycosaminoglycan
  • nucleocytoplasmic O-GlcNAc
  • glycosyltransferase
  • human disease
  • congenital disorders of glycosylation
  • cancer
  • glycan analysis

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Published Papers (17 papers)

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Editorial

Jump to: Research, Review

2 pages, 167 KiB  
Editorial
Special Issue: New Insights into Protein Glycosylation
by Yuuki Kurebayashi and Hideyuki Takeuchi
Molecules 2023, 28(7), 3263; https://doi.org/10.3390/molecules28073263 - 6 Apr 2023
Cited by 1 | Viewed by 1310
Abstract
Protein glycosylation is a general post-translational modification pathway that controls various biological functions including protein trafficking, cell adhesion, and protein-ligand interaction [...] Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)

Research

Jump to: Editorial, Review

11 pages, 2401 KiB  
Article
Characterization of Hyaluronidase 4 Involved in the Catabolism of Chondroitin Sulfate
by Shuhei Yamada and Shuji Mizumoto
Molecules 2022, 27(18), 6103; https://doi.org/10.3390/molecules27186103 - 18 Sep 2022
Cited by 6 | Viewed by 1810
Abstract
Hyaluronidases (HYALs) are endo-beta-N-acetylhexosaminidases that depolymerize not only hyaluronan but also chondroitin sulfate (CS) at the initial step of their catabolism. Although HYAL1 hydrolyzes both CS and HA, HYAL4 is a CS-specific endoglycosidase. The substrate specificity of HYAL4 and identification of [...] Read more.
Hyaluronidases (HYALs) are endo-beta-N-acetylhexosaminidases that depolymerize not only hyaluronan but also chondroitin sulfate (CS) at the initial step of their catabolism. Although HYAL1 hydrolyzes both CS and HA, HYAL4 is a CS-specific endoglycosidase. The substrate specificity of HYAL4 and identification of amino acid residues required for its enzymatic activity have been reported. In this study, we characterized the properties of HYAL4 including the expression levels in various tissues, cellular localization, and effects of its overexpression on intracellular CS catabolism, using cultured cells as well as mouse tissues. Hyal4 mRNA and HYAL4 protein were demonstrated to be ubiquitously expressed in various organs in the mouse. HYAL4 protein was shown to be present both on cell surfaces as well as in lysosomes of rat skeletal muscle myoblasts, L6 cells. Overexpression of HYAL4 in Chinese hamster ovary cells decreased in the total amount of CS, suggesting its involvement in the cellular catabolism of CS. In conclusion, HYAL4 may be widely distributed and play various biological roles, including the intracellular depolymerization of CS. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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13 pages, 2576 KiB  
Article
Complementary Role of GlcNAc6ST2 and GlcNAc6ST3 in Synthesis of CL40-Reactive Sialylated and Sulfated Glycans in the Mouse Pleural Mesothelium
by Yoshiko Takeda-Uchimura, Midori Ikezaki, Tomoya O. Akama, Kaho Nishioka, Yoshito Ihara, Fabrice Allain, Kazuchika Nishitsuji and Kenji Uchimura
Molecules 2022, 27(14), 4543; https://doi.org/10.3390/molecules27144543 - 16 Jul 2022
Cited by 1 | Viewed by 1944
Abstract
Sialyl 6-sulfo Lewis X (6-sulfo sLeX) and its derivative sialyl 6-sulfo N-acetyllactosamine (LacNAc) are sialylated and sulfated glycans of sialomucins found in the high endothelial venules (HEVs) of secondary lymphoid organs. A component of 6-sulfo sLeX present in the [...] Read more.
Sialyl 6-sulfo Lewis X (6-sulfo sLeX) and its derivative sialyl 6-sulfo N-acetyllactosamine (LacNAc) are sialylated and sulfated glycans of sialomucins found in the high endothelial venules (HEVs) of secondary lymphoid organs. A component of 6-sulfo sLeX present in the core 1-extended O-linked glycans detected by the MECA-79 antibody was previously shown to exist in the lymphoid aggregate vasculature and bronchial mucosa of allergic and asthmatic lungs. The components of 6-sulfo sLeX in pulmonary tissues under physiological conditions remain to be analyzed. The CL40 antibody recognizes 6-sulfo sLeX and sialyl 6-sulfo LacNAc in O-linked and N-linked glycans, with absolute requirements for both GlcNAc-6-sulfation and sialylation. Immunostaining of normal mouse lungs with CL40 was performed and analyzed. The contribution of GlcNAc-6-O-sulfotransferases (GlcNAc6STs) to the synthesis of the CL40 epitope in the lungs was also elucidated. Here, we show that the expression of the CL40 epitope was specifically detected in the mesothelin-positive mesothelium of the pulmonary pleura. Moreover, GlcNAc6ST2 (encoded by Chst4) and GlcNAc6ST3 (encoded by Chst5), but not GlcNAc6ST1 (encoded by Chst2) or GlcNAc6ST4 (encoded by Chst7), are required for the synthesis of CL40-positive glycans in the lung mesothelium. Furthermore, neither GlcNAc6ST2 nor GlcNAc6ST3 is sufficient for in vivo expression of the CL40 epitope in the lung mesothelium, as demonstrated by GlcNAc6ST1/3/4 triple-knock-out and GlcNAc6ST1/2/4 triple-knock-out mice. These results indicate that CL40-positive sialylated and sulfated glycans are abundant in the pleural mesothelium and are synthesized complementarily by GlcNAc6ST2 and GlcNAc6ST3, under physiological conditions in mice. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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13 pages, 3397 KiB  
Article
Effects of Roxadustat on Erythropoietin Production in the Rat Body
by Yukiko Yasuoka, Yuichiro Izumi, Takashi Fukuyama, Haruki Omiya, Truyen D. Pham, Hideki Inoue, Tomomi Oshima, Taiga Yamazaki, Takayuki Uematsu, Noritada Kobayashi, Yoshitaka Shimada, Yasushi Nagaba, Tetsuro Yamashita, Masashi Mukoyama, Yuichi Sato, Susan M. Wall, Jeff M. Sands, Noriko Takahashi, Katsumasa Kawahara and Hiroshi Nonoguchi
Molecules 2022, 27(3), 1119; https://doi.org/10.3390/molecules27031119 - 8 Feb 2022
Cited by 3 | Viewed by 2820
Abstract
Anemia is a major complication of chronic renal failure. To treat this anemia, prolylhydroxylase domain enzyme (PHD) inhibitors as well as erythropoiesis-stimulating agents (ESAs) have been used. Although PHD inhibitors rapidly stimulate erythropoietin (Epo) production, the precise sites of Epo production following the [...] Read more.
Anemia is a major complication of chronic renal failure. To treat this anemia, prolylhydroxylase domain enzyme (PHD) inhibitors as well as erythropoiesis-stimulating agents (ESAs) have been used. Although PHD inhibitors rapidly stimulate erythropoietin (Epo) production, the precise sites of Epo production following the administration of these drugs have not been identified. We developed a novel method for the detection of the Epo protein that employs deglycosylation-coupled Western blotting. With protein deglycosylation, tissue Epo contents can be quantified over an extremely wide range. Using this method, we examined the effects of the PHD inhibitor, Roxadustat (ROX), and severe hypoxia on Epo production in various tissues in rats. We observed that ROX increased Epo mRNA expression in both the kidneys and liver. However, Epo protein was detected in the kidneys but not in the liver. Epo protein was also detected in the salivary glands, spleen, epididymis and ovaries. However, both PHD inhibitors (ROX) and severe hypoxia increased the Epo protein abundance only in the kidneys. These data show that, while Epo is produced in many tissues, PHD inhibitors as well as severe hypoxia regulate Epo production only in the kidneys. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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12 pages, 1490 KiB  
Article
A Possible Inhibitory Role of Sialic Acid on MUC1 in Peritoneal Dissemination of Clear Cell-Type Ovarian Cancer Cells
by Yutaka Tamada, Hiroyuki Nomura, Daisuke Aoki and Tatsuro Irimura
Molecules 2021, 26(19), 5962; https://doi.org/10.3390/molecules26195962 - 1 Oct 2021
Cited by 4 | Viewed by 2012
Abstract
The role of sialic acids on MUC1 in peritoneal dissemination of ovarian cancer cells was investigated. A human ovarian carcinoma cell line, ES-2, was transfected with full-length MUC1 containing 22 or 42 tandem repeats. These transfectants were less adherent to monolayers of patient-derived [...] Read more.
The role of sialic acids on MUC1 in peritoneal dissemination of ovarian cancer cells was investigated. A human ovarian carcinoma cell line, ES-2, was transfected with full-length MUC1 containing 22 or 42 tandem repeats. These transfectants were less adherent to monolayers of patient-derived mesothelial cells than ES-2/mock transfectants. When these cells were inoculated into the abdominal cavity of female nude mice, mice that had received the transfectants showed better survival. When the transfectants were mixed with sialidase and injected, the survival was poorer, whereas when they were mixed with N-acetyl-2,3-dehydro-2-deoxyneuraminic acid, a sialidase inhibitor, the survival was significantly prolonged. These behaviors, concerned with peritoneal implantation and dissemination observed in vitro and in vivo, were dependent on the expression of MUC1. Therefore, sialic acid linked to MUC1 in the form, at least in part, of sialyl-T, as shown to be recognized by monoclonal antibody MY.1E12, is responsible for the suppression of adhesion of these cells to mesothelial cells and the suppression of peritoneal implantation and dissemination. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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20 pages, 2471 KiB  
Article
Modulation of the NOTCH1 Pathway by LUNATIC FRINGE Is Dominant over That of MANIC or RADICAL FRINGE
by Florian Pennarubia, Alison V. Nairn, Megumi Takeuchi, Kelley W. Moremen and Robert S. Haltiwanger
Molecules 2021, 26(19), 5942; https://doi.org/10.3390/molecules26195942 - 30 Sep 2021
Cited by 9 | Viewed by 2228
Abstract
Fringes are glycosyltransferases that transfer a GlcNAc to O-fucose residues on Epidermal Growth Factor-like (EGF) repeats. Three Fringes exist in mammals: LUNATIC FRINGE (LFNG), MANIC FRINGE (MFNG), and RADICAL FRINGE (RFNG). Fringe modification of O-fucose on EGF repeats in the NOTCH1 [...] Read more.
Fringes are glycosyltransferases that transfer a GlcNAc to O-fucose residues on Epidermal Growth Factor-like (EGF) repeats. Three Fringes exist in mammals: LUNATIC FRINGE (LFNG), MANIC FRINGE (MFNG), and RADICAL FRINGE (RFNG). Fringe modification of O-fucose on EGF repeats in the NOTCH1 (N1) extracellular domain modulates the activation of N1 signaling. Not all O-fucose residues of N1 are modified by all Fringes; some are modified by one or two Fringes and others not modified at all. The distinct effects on N1 activity depend on which Fringe is expressed in a cell. However, little data is available on the effect that more than one Fringe has on the modification of O-fucose residues and the resulting downstream consequence on Notch activation. Using mass spectral glycoproteomic site mapping and cell-based N1 signaling assays, we compared the effect of co-expression of N1 with one or more Fringes on modification of O-fucose and activation of N1 in three cell lines. Individual expression of each Fringe with N1 in the three cell lines revealed differences in modulation of the Notch pathway dependent on the presence of endogenous Fringes. Despite these cell-based differences, co-expression of several Fringes with N1 demonstrated a dominant effect of LFNG over MFNG or RFNG. MFNG and RFNG appeared to be co-dominant but strongly dependent on the ligands used to activate N1 and on the endogenous expression of Fringes. These results show a hierarchy of Fringe activity and indicate that the effect of MFNG and/or RFNG could be small in the presence of LFNG. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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9 pages, 2318 KiB  
Article
Involvement of DPY19L3 in Myogenic Differentiation of C2C12 Myoblasts
by Kento Mori, Hongkai Sun, Kazuki Miura and Siro Simizu
Molecules 2021, 26(18), 5685; https://doi.org/10.3390/molecules26185685 - 19 Sep 2021
Cited by 5 | Viewed by 3099 | Correction
Abstract
DPY19L3 has been identified as a C-mannosyltransferase for thrombospondin type-1 repeat domain-containing proteins. In this study, we focused on the role of DPY19L3 in the myogenic differentiation of C2C12 mouse myoblast cells. We carried out DPY19L3 gene depletion using the CRISPR/Cas9 system. [...] Read more.
DPY19L3 has been identified as a C-mannosyltransferase for thrombospondin type-1 repeat domain-containing proteins. In this study, we focused on the role of DPY19L3 in the myogenic differentiation of C2C12 mouse myoblast cells. We carried out DPY19L3 gene depletion using the CRISPR/Cas9 system. The result showed that these DPY19L3-knockout cells could not be induced for differentiation. Moreover, the phosphorylation levels of MEK/ERK and p70S6K were suppressed in the DPY19L3-knockout cells compared with that of parent cells, suggesting that the protein(s) that is(are) DPY19L3-mediated C-mannosylated and regulate(s) MEK/ERK or p70S6K signaling is(are) required for the differentiation. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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10 pages, 1971 KiB  
Article
Ribitol in Solution Is an Equilibrium of Asymmetric Conformations
by Shiho Ohno, Noriyoshi Manabe, Takumi Yamaguchi, Jun Uzawa and Yoshiki Yamaguchi
Molecules 2021, 26(18), 5471; https://doi.org/10.3390/molecules26185471 - 8 Sep 2021
Cited by 2 | Viewed by 2415
Abstract
Ribitol (C5H12O5), an acyclic sugar alcohol, is present on mammalian α-dystroglycan as a component of O-mannose glycan. In this study, we examine the conformation and dynamics of ribitol by database analysis, experiments, and computational methods. Database [...] Read more.
Ribitol (C5H12O5), an acyclic sugar alcohol, is present on mammalian α-dystroglycan as a component of O-mannose glycan. In this study, we examine the conformation and dynamics of ribitol by database analysis, experiments, and computational methods. Database analysis reveals that the anti-conformation (180°) is populated at the C3–C4 dihedral angle, while the gauche conformation (±60°) is seen at the C2–C3 dihedral angle. Such conformational asymmetry was born out in a solid-state 13C-NMR spectrum of crystalline ribitol, where C1 and C5 signals are unequal. On the other hand, solution 13C-NMR has identical chemical shifts for C1 and C5. NMR 3J coupling constants and OH exchange rates suggest that ribitol is an equilibrium of conformations, under the influence of hydrogen bonds and/or steric hinderance. Molecular dynamics (MD) simulations allowed us to discuss such a chemically symmetric molecule, pinpointing the presence of asymmetric conformations evidenced by the presence of correlations between C2–C3 and C3–C4 dihedral angles. These findings provide a basis for understanding the dynamic structure of ribitol and the function of ribitol-binding enzymes. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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17 pages, 19408 KiB  
Article
Functional Analysis of the GPI Transamidase Complex by Screening for Amino Acid Mutations in Each Subunit
by Si-Si Liu, Fei Jin, Yi-Shi Liu, Yoshiko Murakami, Yukihiko Sugita, Takayuki Kato, Xiao-Dong Gao, Taroh Kinoshita, Motoyuki Hattori and Morihisa Fujita
Molecules 2021, 26(18), 5462; https://doi.org/10.3390/molecules26185462 - 8 Sep 2021
Cited by 7 | Viewed by 3104
Abstract
Glycosylphosphatidylinositol (GPI) anchor modification is a posttranslational modification of proteins that has been conserved in eukaryotes. The biosynthesis and transfer of GPI to proteins are carried out in the endoplasmic reticulum. Attachment of GPI to proteins is mediated by the GPI-transamidase (GPI-TA) complex, [...] Read more.
Glycosylphosphatidylinositol (GPI) anchor modification is a posttranslational modification of proteins that has been conserved in eukaryotes. The biosynthesis and transfer of GPI to proteins are carried out in the endoplasmic reticulum. Attachment of GPI to proteins is mediated by the GPI-transamidase (GPI-TA) complex, which recognizes and cleaves the C-terminal GPI attachment signal of precursor proteins. Then, GPI is transferred to the newly exposed C-terminus of the proteins. GPI-TA consists of five subunits: PIGK, GPAA1, PIGT, PIGS, and PIGU, and the absence of any subunit leads to the loss of activity. Here, we analyzed functionally important residues of the five subunits of GPI-TA by comparing conserved sequences among homologous proteins. In addition, we optimized the purification method for analyzing the structure of GPI-TA. Using purified GPI-TA, preliminary single particle images were obtained. Our results provide guidance for the structural and functional analysis of GPI-TA. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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10 pages, 2688 KiB  
Article
Effects of Angiotensin II on Erythropoietin Production in the Kidney and Liver
by Yukiko Yasuoka, Yuichiro Izumi, Takashi Fukuyama, Hideki Inoue, Tomomi Oshima, Taiga Yamazaki, Takayuki Uematsu, Noritada Kobayashi, Yoshitaka Shimada, Yasushi Nagaba, Masashi Mukoyama, Yuichi Sato, Jeff M Sands, Katsumasa Kawahara and Hiroshi Nonoguchi
Molecules 2021, 26(17), 5399; https://doi.org/10.3390/molecules26175399 - 5 Sep 2021
Cited by 10 | Viewed by 2904
Abstract
The kidney is a main site of erythropoietin production in the body. We developed a new method for the detection of Epo protein by deglycosylation-coupled Western blotting. Detection of deglycosylated Epo enables the examination of small changes in Epo production. Using this method, [...] Read more.
The kidney is a main site of erythropoietin production in the body. We developed a new method for the detection of Epo protein by deglycosylation-coupled Western blotting. Detection of deglycosylated Epo enables the examination of small changes in Epo production. Using this method, we investigated the effects of angiotensin II (ATII) on Epo production in the kidney. ATII stimulated the plasma Epo concentration; Epo, HIF2α, and PHD2 mRNA expression in nephron segments in the renal cortex and outer medulla; and Epo protein expression in the renal cortex. In situ hybridization and immunohistochemistry revealed that ATII stimulates Epo mRNA and protein expression not only in proximal tubules but also in collecting ducts, especially in intercalated cells. These data support the regulation of Epo production in the kidney by the renin–angiotensin–aldosterone system (RAS). Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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14 pages, 3275 KiB  
Article
Tissue-Specific Regulation of HNK-1 Biosynthesis by Bisecting GlcNAc
by Haruka Kawade, Jyoji Morise, Sushil K. Mishra, Shuta Tsujioka, Shogo Oka and Yasuhiko Kizuka
Molecules 2021, 26(17), 5176; https://doi.org/10.3390/molecules26175176 - 26 Aug 2021
Cited by 4 | Viewed by 3088
Abstract
Human natural killer—1 (HNK-1) is a sulfated glyco-epitope regulating cell adhesion and synaptic functions. HNK-1 and its non-sulfated forms, which are specifically expressed in the brain and the kidney, respectively, are distinctly biosynthesized by two homologous glycosyltransferases: GlcAT-P in the brain and GlcAT-S [...] Read more.
Human natural killer—1 (HNK-1) is a sulfated glyco-epitope regulating cell adhesion and synaptic functions. HNK-1 and its non-sulfated forms, which are specifically expressed in the brain and the kidney, respectively, are distinctly biosynthesized by two homologous glycosyltransferases: GlcAT-P in the brain and GlcAT-S in the kidney. However, it is largely unclear how the activity of these isozymes is regulated in vivo. We recently found that bisecting GlcNAc, a branching sugar in N-glycan, suppresses both GlcAT-P activity and HNK-1 expression in the brain. Here, we observed that the expression of non-sulfated HNK-1 in the kidney is unexpectedly unaltered in mutant mice lacking bisecting GlcNAc. This suggests that the biosynthesis of HNK-1 in the brain and the kidney are differentially regulated by bisecting GlcNAc. Mechanistically, in vitro activity assays demonstrated that bisecting GlcNAc inhibits the activity of GlcAT-P but not that of GlcAT-S. Furthermore, molecular dynamics simulation showed that GlcAT-P binds poorly to bisected N-glycan substrates, whereas GlcAT-S binds similarly to bisected and non-bisected N-glycans. These findings revealed the difference of the highly homologous isozymes for HNK-1 synthesis, highlighting the novel mechanism of the tissue-specific regulation of HNK-1 synthesis by bisecting GlcNAc. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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14 pages, 3450 KiB  
Article
Protein O-Fucosyltransferase 1 Undergoes Interdomain Flexibility in Solution
by Erandi Lira-Navarrete, María Carmen Pallarés, Fabio Castello, Maria J. Ruedas-Rama, Angel Orte, Anabel Lostao and Ramón Hurtado-Guerrero
Molecules 2021, 26(8), 2105; https://doi.org/10.3390/molecules26082105 - 7 Apr 2021
Cited by 5 | Viewed by 2809
Abstract
Protein O-fucosyltransferase 1 (PoFUT1) is a GT-B fold enzyme that fucosylates proteins containing EGF-like repeats. GT-B glycosyltransferases have shown a remarkable grade of plasticity adopting closed and open conformations as a way of tuning their catalytic cycle, a feature that has not [...] Read more.
Protein O-fucosyltransferase 1 (PoFUT1) is a GT-B fold enzyme that fucosylates proteins containing EGF-like repeats. GT-B glycosyltransferases have shown a remarkable grade of plasticity adopting closed and open conformations as a way of tuning their catalytic cycle, a feature that has not been observed for PoFUT1. Here, we analyzed Caenorhabditis elegans PoFUT1 (CePoFUT1) conformational behavior in solution by atomic force microscopy (AFM) and single-molecule fluorescence resonance energy transfer (SMF-FRET). Our results show that this enzyme is very flexible and adopts mainly compact conformations and to a lesser extend a highly dynamic population that oscillates between compact and highly extended conformations. Overall, our experiments illustrate the inherent complexity of CePoFUT1 dynamics, which might play a role during its catalytic cycle. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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11 pages, 2394 KiB  
Communication
Bioinformatics and Functional Analyses Implicate Potential Roles for EOGT and L-fringe in Pancreatic Cancers
by Rashu Barua, Kazuyuki Mizuno, Yuko Tashima, Mitsutaka Ogawa, Hideyuki Takeuchi, Ayumu Taguchi and Tetsuya Okajima
Molecules 2021, 26(4), 882; https://doi.org/10.3390/molecules26040882 - 7 Feb 2021
Cited by 15 | Viewed by 3361
Abstract
Notch signaling receptors, ligands, and their downstream target genes are dysregulated in pancreatic ductal adenocarcinoma (PDAC), suggesting a role of Notch signaling in pancreatic tumor development and progression. However, dysregulation of Notch signaling by post-translational modification of Notch receptors remains poorly understood. Here, [...] Read more.
Notch signaling receptors, ligands, and their downstream target genes are dysregulated in pancreatic ductal adenocarcinoma (PDAC), suggesting a role of Notch signaling in pancreatic tumor development and progression. However, dysregulation of Notch signaling by post-translational modification of Notch receptors remains poorly understood. Here, we analyzed the Notch-modifying glycosyltransferase involved in the regulation of the ligand-dependent Notch signaling pathway. Bioinformatic analysis revealed that the expression of epidermal growth factor (EGF) domain-specific O-linked N-acetylglucosamine (EOGT) and Lunatic fringe (LFNG) positively correlates with a subset of Notch signaling genes in PDAC. The lack of EOGT or LFNG expression inhibited the proliferation and migration of Panc-1 cells, as observed by the inhibition of Notch activation. EOGT expression is significantly increased in the basal subtype, and low expression of both EOGT and LFNG predicts better overall survival in PDAC patients. These results imply potential roles for EOGT- and LFNG-dependent Notch signaling in PDAC. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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Review

Jump to: Editorial, Research

19 pages, 7569 KiB  
Review
Significant Roles of Notch O-Glycosylation in Cancer
by Weiwei Wang, Tetsuya Okajima and Hideyuki Takeuchi
Molecules 2022, 27(6), 1783; https://doi.org/10.3390/molecules27061783 - 9 Mar 2022
Cited by 8 | Viewed by 4146
Abstract
Notch signaling, which was initially identified in Drosophila wing morphogenesis, plays pivotal roles in cell development and differentiation. Optimal Notch pathway activity is essential for normal development and dysregulation of Notch signaling leads to various human diseases, including many types of cancers. In [...] Read more.
Notch signaling, which was initially identified in Drosophila wing morphogenesis, plays pivotal roles in cell development and differentiation. Optimal Notch pathway activity is essential for normal development and dysregulation of Notch signaling leads to various human diseases, including many types of cancers. In hematopoietic cancers, such as T-cell acute lymphoblastic leukemia, Notch plays an oncogenic role, while in acute myeloid leukemia, it has a tumor-suppressive role. In solid tumors, such as hepatocellular carcinoma and medulloblastoma, Notch may have either an oncogenic or tumor-suppressive role, depending on the context. Aberrant expression of Notch receptors or ligands can alter the ligand-dependent Notch signaling and changes in trafficking can lead to ligand-independent signaling. Defects in any of the two signaling pathways can lead to tumorigenesis and tumor progression. Strikingly, O-glycosylation is one such process that modulates ligand–receptor binding and trafficking. Three types of O-linked modifications on the extracellular epidermal growth factor-like (EGF) repeats of Notch receptors are observed, namely O-glucosylation, O-fucosylation, and O-N-acetylglucosamine (GlcNAc) modifications. In addition, O-GalNAc mucin-type O-glycosylation outside the EGF repeats also appears to occur in Notch receptors. In this review, we first briefly summarize the basics of Notch signaling, describe the latest information on O-glycosylation of Notch receptors classified on a structural basis, and finally describe the regulation of Notch signaling by O-glycosylation in cancer. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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14 pages, 1464 KiB  
Review
Biosynthetic Mechanisms and Biological Significance of Glycerol Phosphate-Containing Glycan in Mammals
by Rieko Imae, Hiroshi Manya and Tamao Endo
Molecules 2021, 26(21), 6675; https://doi.org/10.3390/molecules26216675 - 4 Nov 2021
Cited by 2 | Viewed by 2265
Abstract
Bacteria contain glycerol phosphate (GroP)-containing glycans, which are important constituents of cell-surface glycopolymers such as the teichoic acids of Gram-positive bacterial cell walls. These glycopolymers comprising GroP play crucial roles in bacterial physiology and virulence. Recently, the first identification of a GroP-containing glycan [...] Read more.
Bacteria contain glycerol phosphate (GroP)-containing glycans, which are important constituents of cell-surface glycopolymers such as the teichoic acids of Gram-positive bacterial cell walls. These glycopolymers comprising GroP play crucial roles in bacterial physiology and virulence. Recently, the first identification of a GroP-containing glycan in mammals was reported as a variant form of O-mannosyl glycan on α-dystroglycan (α-DG). However, the biological significance of such GroP modification remains largely unknown. In this review, we provide an overview of this new discovery of GroP-containing glycan in mammals and then outline the recent progress in elucidating the biosynthetic mechanisms of GroP-containing glycans on α-DG. In addition, we discuss the potential biological role of GroP modification along with the challenges and prospects for further research. The progress in this newly identified glycan modification will provide insights into the phylogenetic implications of glycan. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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18 pages, 811 KiB  
Review
Polypeptide N-acetylgalactosaminyltransferase-Associated Phenotypes in Mammals
by Kentaro Kato, Lars Hansen and Henrik Clausen
Molecules 2021, 26(18), 5504; https://doi.org/10.3390/molecules26185504 - 10 Sep 2021
Cited by 15 | Viewed by 4843
Abstract
Mucin-type O-glycosylation involves the attachment of glycans to an initial O-linked N-acetylgalactosamine (GalNAc) on serine and threonine residues on proteins. This process in mammals is initiated and regulated by a large family of 20 UDP-GalNAc: polypeptide N-acetylgalactosaminyltransferases (GalNAc-Ts) (EC [...] Read more.
Mucin-type O-glycosylation involves the attachment of glycans to an initial O-linked N-acetylgalactosamine (GalNAc) on serine and threonine residues on proteins. This process in mammals is initiated and regulated by a large family of 20 UDP-GalNAc: polypeptide N-acetylgalactosaminyltransferases (GalNAc-Ts) (EC 2.4.1.41). The enzymes are encoded by a large gene family (GALNTs). Two of these genes, GALNT2 and GALNT3, are known as monogenic autosomal recessive inherited disease genes with well characterized phenotypes, whereas a broad spectrum of phenotypes is associated with the remaining 18 genes. Until recently, the overlapping functionality of the 20 members of the enzyme family has hindered characterizing the specific biological roles of individual enzymes. However, recent evidence suggests that these enzymes do not have full functional redundancy and may serve specific purposes that are found in the different phenotypes described. Here, we summarize the current knowledge of GALNT and associated phenotypes. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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27 pages, 4046 KiB  
Review
Protein C-Mannosylation and C-Mannosyl Tryptophan in Chemical Biology and Medicine
by Shiho Minakata, Shino Manabe, Yoko Inai, Midori Ikezaki, Kazuchika Nishitsuji, Yukishige Ito and Yoshito Ihara
Molecules 2021, 26(17), 5258; https://doi.org/10.3390/molecules26175258 - 30 Aug 2021
Cited by 21 | Viewed by 4458
Abstract
C-Mannosylation is a post-translational modification of proteins in the endoplasmic reticulum. Monomeric α-mannose is attached to specific Trp residues at the first Trp in the Trp-x-x-Trp/Cys (W-x-x-W/C) motif of substrate proteins, by the action of C-mannosyltransferases, DPY19-related gene products. The [...] Read more.
C-Mannosylation is a post-translational modification of proteins in the endoplasmic reticulum. Monomeric α-mannose is attached to specific Trp residues at the first Trp in the Trp-x-x-Trp/Cys (W-x-x-W/C) motif of substrate proteins, by the action of C-mannosyltransferases, DPY19-related gene products. The acceptor substrate proteins are included in the thrombospondin type I repeat (TSR) superfamily, cytokine receptor type I family, and others. Previous studies demonstrated that C-mannosylation plays critical roles in the folding, sorting, and/or secretion of substrate proteins. A C-mannosylation-defective gene mutation was identified in humans as the disease-associated variant affecting a C-mannosylation motif of W-x-x-W of ADAMTSL1, which suggests the involvement of defects in protein C-mannosylation in human diseases such as developmental glaucoma, myopia, and/or retinal defects. On the other hand, monomeric C-mannosyl Trp (C-Man-Trp), a deduced degradation product of C-mannosylated proteins, occurs in cells and extracellular fluids. Several studies showed that the level of C-Man-Trp is upregulated in blood of patients with renal dysfunction, suggesting that the metabolism of C-Man-Trp may be involved in human kidney diseases. Together, protein C-mannosylation is considered to play important roles in the biosynthesis and functions of substrate proteins, and the altered regulation of protein C-manosylation may be involved in the pathophysiology of human diseases. In this review, we consider the biochemical and biomedical knowledge of protein C-mannosylation and C-Man-Trp, and introduce recent studies concerning their significance in biology and medicine. Full article
(This article belongs to the Special Issue New Insights into Protein Glycosylation)
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