Hijacking a biosynthetic pathway yields a glycosyltransferase inhibitor within cells.
Glycosyltransferases are ubiquitous enzymes that catalyze the assembly of glycoconjugates throughout all kingdoms of nature. A long-standing problem is the rational design of probes that can be used to manipulate glycosyltransferase activity in cells and tissues. Here we describe the rational design and synthesis of a nucleotide sugar analog that inhibits, with high potency both in vitro and in cells, the human glycosyltransferase responsible for the reversible post-translational modification of nucleocytoplasmic proteins with O-linked N-acetylglucosamine residues (O-GlcNAc). We show that the enzymes of the hexosamine biosynthetic pathway can transform, both in vitro and in cells, a synthetic carbohydrate precursor into the nucleotide sugar analog. Treatment of cells with the precursor lowers O-GlcNAc in a targeted manner with a single-digit micromolar EC(50). This approach to inhibition of glycosyltransferases should be applicable to other members of this superfamily of enzymes and enable their manipulation in a biological setting.
Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada.
Tracey M
TM
Gloster
Wesley F
WF
Zandberg
Julia E
JE
Heinonen
David L
DL
Shen
Lehua
L
Deng
David J
DJ
Vocadlo
D000118:Acetylglucosaminidase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D053898:Biosynthetic Pathways; D003593:Cytoplasm / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D011499:Protein Processing, Post-Translational / Q000187:drug effects*
D004791:Enzyme Inhibitors; D000118:Acetylglucosaminidase
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