'We report an advanced chemoenzymatic labeling strategy for direct fluorescence detection of O-GlcNAc proteins in gels that facilitates proteomic studies and greatly extend the reach of existing technologies. These new tools also enable the expression and dynamics of O-GlcNAc modifications to be monitored by imaging in cells and tissues. ... More
Comparative methods for analysis of protein covalent modification by electrophilic quinoids formed from xenobiotics.
'Conjugation of biotin and fluorophore tags is useful for assaying covalent protein modification. Oxidative bioactivation of selective estrogen receptor modulators (SERMs) yields reactive quinoid electrophiles that covalently modify proteins, and bioactivation is associated with carcinogenic and chemopreventive effects. Identification of the protein targets of electrophilic metabolites is of general importance ... More
Rapid temporal dynamics of transcription, protein synthesis, and secretion during macrophage activation.
AuthorsEichelbaum K, Krijgsveld J,
Journal
PubMed ID24396086
Macrophages provide the first line of host defense with their capacity to react to an array of cytokines and bacterial components requiring tight regulation of protein expression and secretion to invoke a properly tuned innate immune response. To capture the dynamics of this system, we introduce a novel method combining ... More
A functional RNAi screen links O-GlcNAc modification of ribosomal proteins to stress granule and processing body assembly.
AuthorsOhn T, Kedersha N, Hickman T, Tisdale S, Anderson P,
JournalNat Cell Biol
PubMed ID18794846
Stress granules (SGs) and processing bodies (PBs) are microscopically visible ribonucleoprotein granules that cooperatively regulate the translation and decay of messenger RNA. Using an RNA-mediated interference-based screen, we identify 101 human genes required for SG assembly, 39 genes required for PB assembly, and 31 genes required for coordinate SG and ... More
Identification of structural and functional O-linked N-acetylglucosamine-bearing proteins in Xenopus laevis oocyte.
O-Linked N-acetylglucosaminylation (O-GlcNAcylation) (or O-linked N-acetylglucosamine (O-GlcNAc)) is an abundant and reversible glycosylation type found within the cytosolic and the nuclear compartments. We have described previously the sudden O-GlcNAcylation increase occurring during the Xenopus laevis oocyte G(2)/M transition, and we have demonstrated that the inhibition of O-GlcNAc-transferase (OGT) blocked this ... More
Regulation of calcium/calmodulin-dependent kinase IV by O-GlcNAc modification.
AuthorsDias WB, Cheung WD, Wang Z, Hart GW,
JournalJ Biol Chem
PubMed ID19506079
Similar to phosphorylation, GlcNAcylation (the addition of O-GlcNAc to Ser(Thr) residues on polypeptides) is an abundant, dynamic, and inducible post-translational modification. GlcNAcylated proteins are crucial in regulating virtually all cellular processes, including signaling, cell cycle, and transcription. Here we show that calcium/calmodulin-dependent kinase IV (CaMKIV) is highly GlcNAcylated in vivo. ... More