Structure of transmembrane prolyl 4-hydroxylase reveals unique organization of EF and dioxygenase domains.

Matti Myllykoski, Aleksi Sutinen, M Kristian Koski, Juha P Kallio, Arne Raasakka, Johanna Myllyharju, Rik K Wierenga, Peppi Koivunen

The Journal of biological chemistry 2021 Jan-Jun

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Prolyl 4-hydroxylases (P4Hs) catalyze post-translational hydroxylation of peptidyl proline residues. In addition to collagen P4Hs and hypoxia-inducible factor P4Hs, a third P4H-the poorly characterized endoplasmic reticulum-localized transmembrane prolyl 4-hydroxylase (P4H-TM)-is found in animals. P4H-TM variants are associated with the familiar neurological HIDEA syndrome, but how these variants might contribute to disease is unknown. Here, we explored this question in a structural and functional analysis of soluble human P4H-TM. The crystal structure revealed an EF domain with two Ca2+-binding motifs inserted within the catalytic domain. A substrate-binding groove was formed between the EF domain and the conserved core of the catalytic domain. The proximity of the EF domain to the active site suggests that Ca2+ binding is relevant to the catalytic activity. Functional analysis demonstrated that Ca2+-binding affinity of P4H-TM is within the range of physiological Ca2+ concentration in the endoplasmic reticulum. P4H-TM was found both as a monomer and a dimer in the solution, but the monomer-dimer equilibrium was not regulated by Ca2+. The catalytic site contained bound Fe2+ and N-oxalylglycine, which is an analogue of the cosubstrate 2-oxoglutarate. Comparison with homologous P4H structures complexed with peptide substrates showed that the substrate-interacting residues and the lid structure that folds over the substrate are conserved in P4H-TM, whereas the extensive loop structures that surround the substrate-binding groove, generating a negative surface potential, are different. Analysis of the structure suggests that the HIDEA variants cause loss of P4H-TM function. In conclusion, P4H-TM shares key structural elements with other P4Hs while having a unique EF domain. Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.

Citation

Matti Myllykoski, Aleksi Sutinen, M Kristian Koski, Juha P Kallio, Arne Raasakka, Johanna Myllyharju, Rik K Wierenga, Peppi Koivunen. Structure of transmembrane prolyl 4-hydroxylase reveals unique organization of EF and dioxygenase domains. The Journal of biological chemistry. 2021 Jan-Jun;296:100197