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Posted by / 24-Jul-2020 17:14

Elucidating the role

Our work sheds light on the regulation of CREB through PTMs to modulate neuronal function and delineate the roles of O-Glc NAcylation and phosphorylation in modulating neuronal excitability and neuronal development and metabolism respectively.Altogether, these studies demonstrate that O-Glc NAc modification is a critical mediator of neuronal homeostasis and neurodegeneration.

To help elucidate these mechanisms, our laboratory conducted a genome-wide si RNA screen, which identified several novel regulators of melanogenesis.Due to the lack of a well-defined OGT substrate sequence and structural constraints, OGT is believed to obtain its substrate specificity through its interactome where specific interactors target OGT to specific substrates.In order to identify these interactors, we used CRISPR/Cas9 to generate a novel mouse with a minimally tagged OGT in order to identify the endogenous OGT brain interactome using tandem affinity purification and MS methods.Chemoenzymatic labeling followed by Cu(I)-catalyzed azide-alkyne cycloaddition (Cu AAC) installed a new MS-compatible linker designed for facile purification and release of O-Glc NAcylated proteins for downstream MS analysis.We validated the approach by identifying several established O-Glc NAc sites on the proteins α-crystallin and OGT as well as discovering new, previously unreported sites on both proteins.

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Epidermal melanocytes synthesize melanin within specialized lysosome-related organelles known as melanosomes.

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