Aspartate/asparagine-β-hydroxylase crystal structures reveal an unexpected epidermal growth factor-like domain substrate disulfide pattern
Pfeffer, Inga, Brewitz, Lennart, Krojer, Tobias, Jensen, Sacha A., Kochan, Grazyna T., Kershaw, Nadia J., Hewitson, Kirsty S., McNeill, Luke A., Kramer, Holger, Münzel, Martin, Hopkinson, Richard J., Oppermann, Udo, Handford, Penny A., McDonough, Michael A., and Schofield, Christopher J. (2019) Aspartate/asparagine-β-hydroxylase crystal structures reveal an unexpected epidermal growth factor-like domain substrate disulfide pattern. Nature Communications, 10. 4910.
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Abstract
AspH is an endoplasmic reticulum (ER) membrane-anchored 2-oxoglutarate oxygenase whose C-terminal oxygenase and tetratricopeptide repeat (TPR) domains present in the ER lumen. AspH catalyses hydroxylation of asparaginyl- and aspartyl-residues in epidermal growth factor-like domains (EGFDs). Here we report crystal structures of human AspH, with and without substrate, that reveal substantial conformational changes of the oxygenase and TPR domains during substrate binding. Fe(II)-binding by AspH is unusual, employing only two Fe(II)-binding ligands (His679/His725). Most EGFD structures adopt an established fold with a conserved Cys1–3, 2–4, 5–6 disulfide bonding pattern; an unexpected Cys3–4 disulfide bonding pattern is observed in AspH-EGFD substrate complexes, the catalytic relevance of which is supported by studies involving stable cyclic peptide substrate analogues and by effects of Ca(II) ions on activity. The results have implications for EGFD disulfide pattern processing in the ER and will enable medicinal chemistry efforts targeting human 2OG oxygenases.
| Item ID: | 60806 |
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| Item Type: | Article (Research - C1) |
| ISSN: | 2041-1723 |
| Copyright Information: | © The Author(s) 2019. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
| Funders: | Wellcome Trust, Cancer Research UK, Biotechnological and Biological Research Council, European Union’s Seventh Framework Programme (FP7), St Edmund Hall Oxford (SEHO), Deutsche Forschungsgemeinschaft (DFG), Arthritis Research UK (AR) |
| Projects and Grants: | FP7 grant 298603, SEHO William R. Miller Junior Research Fellowship, DFG fellowship BR 5486/2-1, AR grant 20785, AR grant 20522 |
| Date Deposited: | 05 Nov 2019 04:30 |
| FoR Codes: | 31 BIOLOGICAL SCIENCES > 3101 Biochemistry and cell biology > 310112 Structural biology (incl. macromolecular modelling) @ 40% 31 BIOLOGICAL SCIENCES > 3101 Biochemistry and cell biology > 310105 Cellular interactions (incl. adhesion, matrix, cell wall) @ 20% 31 BIOLOGICAL SCIENCES > 3101 Biochemistry and cell biology > 310108 Protein trafficking @ 40% |
| SEO Codes: | 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100% |
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