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Aspartate residues far from the active site drive O-GlcNAc transferase substrate selection

ABSTRACT: Here we use protein microarray technology and proteome-wide glycosylation profiling to show that conserved aspartate residues in the tetratricopeptide repeat (TPR) lumen of OGT drive substrate selection. Changing these residues to alanines alters substrate selectivity and unexpectedly increases rates of protein glycosylation. Our findings support a model where sites of glycosylation for many OGT substrates are determined by TPR domain contacts to substrate side chains five to fifteen residues Cterminal to the glycosite. In addition to guiding design of inhibitors that target OGT’s TPR domain, this information will inform efforts to engineer substrates to explore biological functions.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Christina Woo

LAB HEAD: Christina Woo and Suzanne Walker

PROVIDER: PXD014731 | Pride | 2019-07-26

REPOSITORIES: Pride

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Aspartate Residues Far from the Active Site Drive O-GlcNAc Transferase Substrate Selection.

Joiner Cassandra M CM Levine Zebulon G ZG Aonbangkhen Chanat C Woo Christina M CM Walker Suzanne S

Journal of the American Chemical Society 20190807 33

O-GlcNAc is an abundant post-translational modification found on nuclear and cytoplasmic proteins in all metazoans. This modification regulates a wide variety of cellular processes, and elevated O-GlcNAc levels have been implicated in cancer progression. A single essential enzyme, O-GlcNAc transferase (OGT), is responsible for all nucleocytoplasmic O-GlcNAcylation. Understanding how this enzyme chooses its substrates is critical for understanding, and potentially manipulating, its functions. Her ...[more]

PMID: 31373491

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