Koski, M., Anantharajan, J., Kursula, P., Dhavala, P., Murthy, A., Bergmann, U., Myllyharju, J., Wierenga, R. (2017) Assembly of the elongated collagen prolyl 4-hydroxylase α 2 β 2 heterotetramer around a central α 2 dimer. Biochemical Journal, 474 (5), 751-769. doi:10.1042/BCJ20161000
Assembly of the elongated collagen prolyl 4-hydroxylase α2β2 heterotetramer around a central α2 dimer
|Author:||Koski, M. Kristian1; Anantharajan, Jothi1; Kursula, Petri1,2;|
1Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, PO Box 5400, FIN-90014 Oulu, Finland
2Department of Biomedicine, University of Bergen, PO Box 7804, N-5009 Bergen, Norway
3Oulu Center for Cell-Matrix Research, Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, PO Box 5400, FIN-90014 Oulu, Finland
|Online Access:||PDF Full Text (PDF, 1.7 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe201902205810
|Publish Date:|| 2019-02-20
Collagen prolyl 4-hydroxylase (C-P4H), an α2β2 heterotetramer, is a crucial enzyme for collagen synthesis. The α-subunit consists of an N-terminal dimerization domain, a central peptide substrate-binding (PSB) domain, and a C-terminal catalytic (CAT) domain. The β-subunit [also known as protein disulfide isomerase (PDI)] acts as a chaperone, stabilizing the functional conformation of C-P4H. C-P4H has been studied for decades, but its structure has remained elusive. Here, we present a three-dimensional small-angle X-ray scattering model of the entire human C-P4H-I heterotetramer. C-P4H is an elongated, bilobal, symmetric molecule with a length of 290 Å. The dimerization domains from the two α-subunits form a protein–protein dimer interface, assembled around the central antiparallel coiled-coil interface of their N-terminal α-helices. This region forms a thin waist in the bilobal tetramer. The two PSB/CAT units, each complexed with a PDI/β-subunit, form two bulky lobes pointing outward from this waist region, such that the PDI/β-subunits locate at the far ends of the βααβ complex. The PDI/β-subunit interacts extensively with the CAT domain. The asymmetric shape of two truncated C-P4H-I variants, also characterized in the present study, agrees with this assembly. Furthermore, data from these truncated variants show that dimerization between the α-subunits has an important role in achieving the correct PSB–CAT assembly competent for catalytic activity. Kinetic assays with various proline-rich peptide substrates and inhibitors suggest that, in the competent assembly, the PSB domain binds to the procollagen substrate downstream from the CAT domain.
|Pages:||751 - 769|
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
1182 Biochemistry, cell and molecular biology
This work was supported by the EU-BIOXHIT grant to R.K.W. and by the Academy of Finland grants to R.K.W. , to M.K.K. , to J.M. [200471, 202469, and the Center of Excellence Grant 251314], and to P.K. [108423, 214317, 124241, and 218045]. We also acknowledge the Biocenter Finland support to the Biocenter Oulu Protein Crystallography Core Facility co-ordinated by R.K.W. The work was also funded by a fellowship from the CIMO organization to J.A., grants from the Sigrid Jusélius Foundation to M.K.K./R.K.W., P.K., and J.M., a Jane and Aatos Erkko Foundation grant to J.M., and funding from the Research and Science Foundation of the City of Hamburg (P.K.). The research leading to these results has also received funding from the European Community’s Seventh Framework Programme [FP7/2007-2013] under BioStruct-X [grant agreement no. 283570].
|EU Grant Number:||
(283570) BIOSTRUCT-X - Transnational access and enhancement of integrated Biological Structure determination at synchrotron X-ray radiation facilities
|Academy of Finland Grant Number:||
251314 (Academy of Finland Funding decision)
214317 (Academy of Finland Funding decision)
124241 (Academy of Finland Funding decision)
218045 (Academy of Finland Funding decision)
Copyright 2016 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.