Substrate and product analogues as human O-GlcNAc transferase inhibitors

Dorfmueller, Helge C.; Borodkin, Vladimir S.; Blair, D.E.; Pathak, Shalini; Navrátilová, Iva; Aalten, Daan M. F. van

doi:10.1007/s00726-010-0688-y

Cited by 65 publications

(54 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…O-GlcNAc levels can be directly or indirectly altered in vitro and in vivo by modulating the activity or expression of OGT (Gross et al 2005(Gross et al , 2011Dorfmueller et al 2011), O-GlcNAcase (Dong and Hart 1994;Haltiwanger et al 1998;Liu et al 2002;Dorfmueller et al 2006;Whitworth et al 2007;Macauley et al 2008;Knapp et al 2007;Beer et al 1990;Yuzwa et al 2008;Konrad et al 2002;Laczy et al 2010), or other key enzymes in the HBP (Buse et al 2002;McClain 2002;Patti et al 1999;Ross et al 2000;Marshall et al 1991;Traxinger and Marshall 1991). Techniques such as over expression of OGT (Zachara et al 2004b), OGlcNAcase (Slawson et al 2005), and GFAT (Chen et al 1997;Marshall et al 2004;James et al 2000) by both transient transfection and viral transduction have been successful, as has reducing the expression of these enzymes using RNA interference (Zachara et al 2004b;Ngoh et al 2009a;Hsieh et al 2012).…”

Section: In Vitro and In Vivo Modulation Of O-glcnac Levelsmentioning

confidence: 99%

Section: Murine Models and The Ogt F/ymer-cre-2a-gfp Cell Linementioning

confidence: 99%

“…A more recent study has focused on synthesizing novel OGT inhibitors with structural analogues to UDPGlcNAc/UDP (Dorfmueller et al 2011). While the synthesized compounds inhibit OGT in the micromolar range, it should be noted that they are inactive when used on living cells (Dorfmueller et al 2011). Another recent study developed a synthetic carbohydrate precursor, 2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxy-5-thio-α-d-glucopyranose, which can be successfully converted by the cell to produce UDP5SGlcNAc in cell culture and in vitro models (Gloster et al 2011).…”

Section: Murine Models and The Ogt F/ymer-cre-2a-gfp Cell Linementioning

confidence: 99%

See 2 more Smart Citations

Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis

Groves

Lee

Yildirir

et al. 2013

Cell Stress and Chaperones

117

112

View full text Add to dashboard Cite

O-linked N-acetyl-β-D-glucosamine (O-GlcNAc) is a ubiquitous and dynamic post-translational modification known to modify over 3,000 nuclear, cytoplasmic, and mitochondrial eukaryotic proteins. Addition of O-GlcNAc to proteins is catalyzed by the O-GlcNAc transferase and is removed by a neutral-N-acetyl-β-glucosaminidase (OGlcNAcase). O-GlcNAc is thought to regulate proteins in a manner analogous to protein phosphorylation, and the cycling of this carbohydrate modification regulates many cellular functions such as the cellular stress response. Diverse forms of cellular stress and tissue injury result in enhanced O-GlcNAc modification, or O-GlcNAcylation, of numerous intracellular proteins. Stress-induced OGlcNAcylation appears to promote cell/tissue survival by regulating a multitude of biological processes including: the phosphoinositide 3-kinase/Akt pathway, heat shock protein expression, calcium homeostasis, levels of reactive oxygen species, ER stress, protein stability, mitochondrial dynamics, and inflammation. Here, we will discuss the regulation of these processes by O-GlcNAc and the impact of such regulation on survival in models of ischemia reperfusion injury and trauma hemorrhage. We will also discuss the misregulation of O-GlcNAc in diseases commonly associated with the stress response, namely Alzheimer's and Parkinson's diseases. Finally, we will highlight recent advancements in the tools and technologies used to study the O-GlcNAc modification.

show abstract

Section: In Vitro and In Vivo Modulation Of O-glcnac Levelsmentioning

confidence: 99%

Section: Murine Models and The Ogt F/ymer-cre-2a-gfp Cell Linementioning

confidence: 99%

Section: Murine Models and The Ogt F/ymer-cre-2a-gfp Cell Linementioning

confidence: 99%

See 1 more Smart Citation

Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis

Groves

Lee

Yildirir

et al. 2013

Cell Stress and Chaperones

117

112

View full text Add to dashboard Cite

show abstract

“…[40] . Van Aalten 小组合成了 UDP-GlcNAc 的 S-1-磷酸盐类似物(3, 图 9)和 UDP 的膦酸盐类似物(4, 图 9)作为 OGT 的抑制剂, 以模型肽段代替蛋白质作为底物进行活性测定时, 发现化合物 2～4 和 UDP 的 IC 50 均在微摩尔级别(1.8～93 μmol/L) [41] . 可见, 使用蛋白质作为底物和使用模型肽段作为底物进行活性测试的结果差别很大, 因为在体内 OGT 的天然底物为蛋白质, 所以使用蛋白质作为底物更加科学.…”

Section: 使用放射性标记的糖基供体底物unclassified

O-GlcNAc Transferase and Its Inhibitors

Wang¹

2013

Acta Chim. Sinica

View full text Add to dashboard Cite

O-GlcNAc transferase (OGT), one of essential mammalian enzymes, catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine (UDP-GlcNAc) to hydroxyl groups of serines and threonines (Ser/Thr) in proteins. O-GlcNAcylation is widely located in cytoplasm and nucleus. This kind of protein post-translational modification is involved in the regulation of many cellular signaling pathways and closely associated with the occurrences and developments of numerous critical illnesses. In animals, OGT is encoded by a single highly conserved gene. However, human OGT includes three different isoforms at least: nucleocytoplasmic OGT (ncOGT), mitochondrial OGT (mOGT) and short OGT (sOGT). All of the isoforms comprise two distinct regions: a multidomain catalytic region and an N-terminal region with different numbers of tetratricopeptide repeats (TPRs), which is closely related to their subcellular localizations. The structures of the catalytic region and TPRs were reported and it was supported that OGT uses an ordered sequential "bi-bi" mechanism though the details of catalytic mechanism were not completely understood. To discover the inhibitors of OGT, several OGT activity assay methods were developed. The conventional method uses a radiolabeled glycosyl donor substrate such as UDP-[3 H]-GlcNAc or UDP-[ 14 C]-GlcNAc so that it is not suitable for in vivo test or rapid analysis. O-GlcNAc antibody-based western blot was performed to measure the activity of OGT in vivo, but it is infeasible to determine low abundance protein with single O-GlcNAc. Three high throughput activity assays were reported: ligand displacement OGT assay, protease-protection assay strategy and azido-enzyme-linked immunosorbent assay (azido-ELISA). The first two methods have especially been used to find new OGT inhibitors through screening compound libraries, but the results have to be verified by the conventional method. To date, OGT inhibitor activity of several candidates was evaluated and as found, two of them {2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxy-5-thio-α-D-glucopyranose and 4-methoxyphenyl 6-acetyl-2-oxobenzo[d]-oxazole-3(2H)-carboxylate} worked effectively in vivo and were valuable for understanding the functional roles of OGT. Nevertheless, there are still important questions to be answered in the research of OGT: (1) What is the general base that catalyzes O-GlcNAcylation reaction? (2) How can OGT recognize so many protein substances with the preservation of substance specificity? (3) How can we develop robust high-throughput OGT assays and isoform-specific OGT inhibitors? In conclusion, there is a long way to go for comprehensive understanding of OGT.

show abstract

“…For example, the short OGA exhibits comparative resistance to PugNAc and NAG-thiazoline, but is very sensitive to alpha-GlcNAc thiolsulfonate (Zachara & Hart, 2006). Inhibition of OGT and OGA represents an area of great interest on O-GlcNAcylation research, which is evident from the increasing number of studies addressing the enzymes molecular mechanisms for the addition and removal of O-GlcNAc (Borodkin & van Aalten, 2010;Dorfmueller et al, , 2011Gloster et al, 2011;Gloster & Vocadlo, 2010;Lameira et al, 2011;Lazarus et al, 2011;Li et al, 2011;Macauley & Vocadlo, 2010;MartinezFleites et al, 2010).…”

Section: Introductionmentioning

confidence: 99%