Asparaginyl hydroxylation of the notch ankyrin repeat domain by factor inhibiting hypoxia-inducible factor

Mathew L. Coleman, Michael A. McDonough, Kirsty S. Hewitson, Charlotte Coles, Jasmin Mecinović, Mariola Edelmann, Kristina M. Cook, Matthew E. Cockman, David E. Lancaster, Benedikt M. Kessler, Neil J. Oldham, Peter J. Ratcliffe, Christopher J. Schofield*

*Corresponding author for this work

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Abstract

The stability and activity of hypoxia-inducible factor (HIF) are regulated by the post-translational hydroxylation of specific prolyl and asparaginyl residues. We show that the HIF asparaginyl hydroxylase, factor inhibiting HIF (FIH), also catalyzes hydroxylation of highly conserved asparaginyl residues within ankyrin repeat (AR) domains (ARDs) of endogenous Notch receptors. AR hydroxylation decreases the extent of ARD binding to FIH while not affecting signaling through the canonical Notch pathway. ARD proteins were found to efficiently compete with HIF for FIH-dependent hydroxylation. Crystallographic analyses of the hydroxylated Notch ARD (2.35 Å) and of Notch peptides bound to FIH (2.4-2.6Å) reveal the stereochemistry of hydroxylation on the AR and imply that significant conformational changes are required in the ARD fold in order to enable hydroxylation at the FIH active site. We propose that ARD proteins function as natural inhibitors of FIH and that the hydroxylation status of these proteins provides another oxygen-dependent interface that modulates HIF signaling.

Original languageEnglish
JournalJournal of Biological Chemistry
Volume282
Issue number33
Pages (from-to)24027-24038
Number of pages12
ISSN0021-9258
DOIs
Publication statusPublished - 17. Aug 2007
Externally publishedYes

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Coleman, M. L., McDonough, M. A., Hewitson, K. S., Coles, C., Mecinović, J., Edelmann, M., Cook, K. M., Cockman, M. E., Lancaster, D. E., Kessler, B. M., Oldham, N. J., Ratcliffe, P. J., & Schofield, C. J. (2007). Asparaginyl hydroxylation of the notch ankyrin repeat domain by factor inhibiting hypoxia-inducible factor. Journal of Biological Chemistry, 282(33), 24027-24038. https://doi.org/10.1074/jbc.M704102200