Protein-specific imaging of posttranslational modifications

Lin, Wei; Gao, Ling; Chen, Xing

doi:10.1016/j.cbpa.2015.07.020

Cited by 19 publications

(12 citation statements)

References 53 publications

(53 reference statements)

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“…Post-translational modifications (PTMs) play a significant role in the regulation of protein structure, enzymatic activity, stability or degradation, subcellular localization, protein-protein interactions, and diverse cell signaling (Azevedo and Saiardi 2016; Drazic, et al 2016; Lin, et al 2015; Rodriguez 2014). Many amino acid side chains such as cysteine (C), serine (S), threonine (T), and tyrosine (Y) are post-translationally modified; however, the amino acid lysine (K) is targeted by an extremely high number of PTMs including methylation, ubiquitination, sumoylation, and acetylation.…”

Section: A Role For Posttranslational Modification In Tr Localizationmentioning

confidence: 99%

Thyroid hormone receptor localization in target tissues

Anyetei‐Anum¹,

Roggero²,

Allison³

2018

Journal of Endocrinology

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The thyroid hormone receptors, TRα1, TRβ1 and other subtypes, are members of the nuclear receptor superfamily that mediate the action of thyroid hormone signaling in numerous tissues to regulate important physiological and developmental processes. Their most well-characterized role is as ligand-dependent transcription factors; TRs bind thyroid hormone response elements in the presence or absence of thyroid hormone to facilitate the expression of target genes. Although primarily residing in the nucleus, TRα1 and TRβ1 shuttle rapidly between the nucleus and cytoplasm. We have identified multiple nuclear localization signals and nuclear export signals within TRα1 and TRβ1 that interact with importins and exportins, respectively, to mediate translocation across the nuclear envelope. More recently, enigmatic cytoplasmic functions have been ascribed to other TR subtypes, expanding the diversity of the cellular response to thyroid hormone. By integrating data on localization signal motifs, this review provides an overview of the complex interplay between TR's dynamic transport pathways and thyroid hormone signaling activities. We examine the variation in TR subtype response to thyroid hormone signaling, and what is currently known about regulation of the variety of tissue-specific localization patterns, including targeting to the nucleus, the mitochondria and the inner surface of the plasma membrane.

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Section: A Role For Posttranslational Modification In Tr Localizationmentioning

confidence: 99%

Thyroid hormone receptor localization in target tissues

Anyetei‐Anum¹,

Roggero²,

Allison³

2018

Journal of Endocrinology

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“…Alternatively, we sought to develop a general strategy for protein‐specific imaging of O‐GlcNAcylation in single cells, which exploits the spatial proximity of the O ‐GlcNAc moiety and the attaching protein (Figure ). Our strategy is inspired by and shares a similar design principle with the previously reported methods for protein‐specific imaging of sialylation and phosphorylation of cell‐surface receptors . The protein of interest is specifically tagged with the genetically encoded enhanced green fluorescence protein (EGFP); a second fluorophore, named the O ‐GlcNAc tag, is conjugated onto the O ‐GlcNAc moiety in a pan‐specific manner on various O‐GlcNAcylated proteins in the cells.…”

Section: Figurementioning

confidence: 99%

“…Our strategyi si nspired by and shares as imilar design principle with the previously reported methods for protein-specific imaging of sialylation and phosphorylation of cell-surface receptors. [15][16][17][18][19] The protein of interesti ss pecificallyt agged with the genetically encoded enhanced green fluorescence protein (EGFP);asecond fluorophore, named the O-GlcNAc tag, is conjugatedo nto the O-GlcNAc moiety in ap an-specific mannero nv arious O-GlcNAcylated proteins in the cells. EGFP and the O-GlcNAct ag form aF RET donor-acceptor pair.A sF RET requires the donor-acceptor distance to be within 10 nm, and the FRET efficiency sharplyd ecreases as the sixth powero ft he distance, efficient FRET preferentially occurs intramolecularly when the protein of interesti sO -GlcNAcylated.…”

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confidence: 99%

Protein‐Specific Imaging of O‐GlcNAcylation in Single Cells

2015

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Thousands of intracellular proteins are post-translationally modified with O-GlcNAc, and O-GlcNAcylation impacts the function of modified proteins and mediates diverse biological processes. However, the ubiquity of this important glycosylation makes it highly challenging to probe the O-GlcNAcylation state of a specific protein at the cellular level. Herein, we report the development of a FLIM-FRET-based strategy, which exploits the spatial proximity of the O-GlcNAc moiety and the attaching protein, for protein-specific imaging of O-GlcNAcylation in single cells. We demonstrated this strategy by imaging the O-GlcNAcylation state of tau and β-catenin inside the cells. Furthermore, the changes in tau O-GlcNAcylation were monitored when the overall cellular O-GlcNAc was pharmacologically altered by using the OGT and OGA inhibitors. We envision that the FLIM-FRET strategy will be broadly applicable to probe the O-GlcNAcylation state of various proteins in the cells.

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“…PLA is versatile and has been widely used to visualize protein-protein interactions 103,104 and post-translational proteoforms, the latter through the use of one antibody targeting the protein and the other targeting the PTM. 105 Beyond antibody-mediated labeling, PLA can be coupled with other protein and PTM labeling strategies, provided that these strategies enable linkage of the PLA oligonucleotides to moieties of interest.…”

Section: Pla-based Proteoform Detectionmentioning

confidence: 99%