Study of cross talk between phosphatases and OGA on a ZO-3-derived peptide

Sharif, Suhela; Shi, Jie; Ruijtenbeek, Rob; Pieters, Roland J.

doi:10.1007/s00726-019-02699-1

Cited by 5 publications

(6 citation statements)

References 17 publications

(15 reference statements)

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“…As a useful tool, peptide arrays also benefit O-GlcNAc research . For example, peptide microarrays of different formats have been utilized for (1) the study of enzymatic activity, substrates, and recognition specificity of OGT, , (2) the evaluation of activity and substrate specificity of OGA, (3) screening of inhibitors, and (4) the investigation of the cross-talk between O-GlcNAcylation and phosphorylation. , …”

Section: Large-scale Characterization Of O-glcnac Proteinsmentioning

confidence: 99%

“…492 For example, peptide microarrays of different formats have been utilized for (1) the study of enzymatic activity, substrates, and recognition specificity of OGT, 184,267 (2) the evaluation of activity and substrate specificity of OGA, 337 (3) screening of inhibitors, 493 and (4) the investigation of the cross-talk between O-GlcNAcylation and phosphorylation. 494,495 As a high-throughput technique, protein/peptide microarrays have the capacity to maximize the number of simultaneously analyzed samples or the number of simultaneous measurements on a single sample. Although the arrays cannot be used to generate straightforward sequence information, they have their own valuable uses.…”

Section: Protein/peptide Microarraysmentioning

confidence: 99%

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Analytical and Biochemical Perspectives of Protein O-GlcNAcylation

2021

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Protein O-linked β-N-acetylglucosamine (O-GlcNAc) modification (O-GlcNAcylation) is a unique monosaccharide modification discovered in the early 1980s. With the technological advances in the past several decades, great progress has been made to reveal the biochemistry of O-GlcNAcylation, the substrates of O-GlcNAcylation, and the functional importance of protein O-GlcNAcylation. As a nutrient sensor, protein O-GlcNAcylation plays important roles in almost all biochemical processes examined. Although the functional importance of O-GlcNAcylation of proteins has been extensively reviewed previously, the chemical and biochemical aspects have not been fully addressed. In this review, by critically evaluating key publications in the past 35 years, we aim to provide a comprehensive understanding of this important post-translational modification (PTM) from analytical and biochemical perspectives. Specifically, we will cover (1) multiple analytical advances in the characterization of O-GlcNAc cycling components (i.e., the substrate donor UDP-GlcNAc, the two key enzymes O-GlcNAc transferase and O-GlcNAcase, and O-GlcNAc substrate proteins), (2) the biochemical characterization of the enzymes with a variety of chemical tools, and (3) exploration of O-GlcNAc cycling and its modulating chemicals as potential biomarkers and therapeutic drugs for diseases. Last but not least, we will discuss the challenges and possible solutions for basic and translational research of protein O-GlcNAcylation in the future.

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Section: Large-scale Characterization Of O-glcnac Proteinsmentioning

confidence: 99%

Section: Protein/peptide Microarraysmentioning

confidence: 99%

Analytical and Biochemical Perspectives of Protein O-GlcNAcylation

2021

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“…This approach does not reveal the specific post-translationally modified amino acid residues linked to PTM crosstalk, although thousands of crosstalk sites are identified. Other work has systematically looked at the co-occurrence of PTMs on synthetic peptides from within proteins with the view to predict or even rule out sequence motifs that may be involved in PTM crosstalk. − However, this does not reflect proteins in their physiological environment. Thus, an enhanced methodology for detecting PTM crosstalk in vivo is needed.…”

Section: Introductionmentioning

confidence: 99%

FAIMS Enhances the Detection of PTM Crosstalk Sites

et al. 2022

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Protein post-translational modifications (PTMs) enable cells to rapidly change in response to biological stimuli. With hundreds of different PTMs, understanding these control mechanisms is complex. To date, efforts have focused on investigating the effect of a single PTM on protein function. Yet, many proteins contain multiple PTMs. Moreover, one PTM can alter the prevalence of another, a phenomenon termed PTM crosstalk. Understanding PTM crosstalk is critical; however, its detection is challenging since PTMs occur substoichiometrically. Here, we develop an enrichment-free, label-free proteomics method that utilizes high-field asymmetric ion mobility spectrometry (FAIMS) to enhance the detection of PTM crosstalk. We show that by searching for multiple combinations of dynamic PTMs on peptide sequences, a 6-fold increase in candidate PTM crosstalk sites is identified compared with that of standard liquid chromatography-tandem mass spectrometry (LC-MS/MS) workflows. Additionally, by cycling through FAIMS compensation voltages within a single LC-FAIMS-MS/MS run, we show that our LC-FAIMS-MS/MS workflow can increase multi-PTM-containing peptide identifications without additional increases in run times. With 159 novel candidate crosstalk sites identified, we envisage LC-FAIMS-MS/MS to play an important role in expanding the repertoire of multi-PTM identifications. Moreover, it is only by detecting PTM crosstalk that we can “see” the full picture of how proteins are regulated.

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“…This promiscuous substrate selectivity is likely in-part due to the predominant interaction of OGA with amide backbone atoms within the bound glycopeptide. As seen elsewhere [23], the rate of OGA catalyzed hydrolysis of an O-GlcNAcylated serine was shown to be reduced by half in the presence of an adjacent phosphotyrosine on a ZO3-derived model peptide (RESSYDIY(p)RVPSS(g)QS) [25]. The ability for PTMs to influence OGA catalyzed removal merits further investigation.…”

Section: Ogt Structure and Polypeptide Acceptor Specificitymentioning

confidence: 61%

Molecular mechanisms regulating O-linked N-acetylglucosamine (O-GlcNAc)–processing enzymes

King

Males

Davies

et al. 2019

Current Opinion in Chemical Biology

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The post-translational modification of proteins by O-linked N-acetylglucosamine (O-GlcNAc) dynamically programs cellular physiology to maintain homeostasis and tailor biochemical pathways to meet context-dependant cellular needs. Despite diverse roles for O-GlcNAc, only two enzymes act antagonistically to govern its cycling; O-GlcNAc transferase (OGT) installs the monosaccharide on target proteins and O-GlcNAc hydrolase (OGA) removes it. Recent literature has exposed a network of mechanisms regulating these two enzymes to choreograph global, and target-specific, O-GlcNAc cycling in response to cellular stress and nutrient availability. Herein, we amalgamate these emerging mechanisms from a structural and molecular perspective to explore how the cell exerts fine control to regulate O-GlcNAcylation of diverse proteins in a selective fashion. Papers of particular interest, published within the period of review, have been highlighted as: *of special interest ** of outstanding interest **[3] An important paper in the field. The authors provide unambiguous evidence for mutations in ogt implicated in XLID influencing OGT function. Despite largely normal global O-GlcNAc levels, significant changes in gene transcription are observed. Furthermore, a compensatory mechanism to maintain global O-GlcNAc levels involving OGT-mSin3A-HDAC1 dependant transcriptional repression at the OGA promoter is proposed. *[6] The authors develop a systematic platform for glycosylation sequence characterization and optimization and use this method to characterize OGT sequence preference. This analysis revealed a surprising preference for an aromatic residue at the-4 subsite. *[10] An asparagine ladder in the TPR domains mediates OGT interaction with protein substrates. * [12] New chemical biology tools to identify features of OGT responsible for substrate targeting. This method uncovered a patch of residues lining the inner surface of the N-terminal domain that contribute to interaction with multiple substrates. **[13] A nice structural study defines the effect of XLID-associated OGT mutation L254F as distorting the TPR helix. * [27] mOGT was shown to be catalytically active in vivo and plays an important role in supporting mitochondrial structure and function. **[28] A fundamental advance in discovery of the OGT NLS and the effects of S389 O-GlcNAcylation and interaction with importin α5 for nuclear import. *[35] The authors demonstrate that OGT is enriched in the post-synaptic density of excitatory neuronal synapses whereby it regulates synapse maturation. *[37] Upon glucagon-induced calcium signalling CamKII phosphorylates OGT at S20, which induces O-GlcNAc modification of Ulk proteins through potentiation of AMPKdependant phosphorylation. **[38] Chk1 phosphorylates OGT S20, and this modification regulates OGT localization to the mid-body and regulation of vimentin bridge formation and severing during mitosis. *[39] LSD2 was shown to act as an E3 ligase to ubiquitinylate OGT and thereby facilitate its UPS mediated degradation. **[43] Circad...

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Study of cross talk between phosphatases and OGA on a ZO-3-derived peptide

Cited by 5 publications

References 17 publications

Analytical and Biochemical Perspectives of Protein O-GlcNAcylation

Analytical and Biochemical Perspectives of Protein O-GlcNAcylation

FAIMS Enhances the Detection of PTM Crosstalk Sites

Molecular mechanisms regulating O-linked N-acetylglucosamine (O-GlcNAc)–processing enzymes

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