Sridhar, S., Schmitz, W., Hiltunen, J. K., Venkatesan, R., Bergmann, U., Kiema, T.-R., & Wierenga, R. K. (2020). Crystallographic binding studies of rat peroxisomal multifunctional enzyme type 1 with 3-ketodecanoyl-CoA: capturing active and inactive states of its hydratase and dehydrogenase catalytic sites. Acta Crystallographica Section D Structural Biology, 76(12), 1256–1269. https://doi.org/10.1107/s2059798320013819
Crystallographic binding studies of rat peroxisomal multifunctional enzyme, type-1 (MFE1) with 3-ketodecanoyl-CoA : capturing active and inactive states of its hydratase and dehydrogenase catalytic sites
|Author:||Sridhar, Shruthi1; Schmitz, Werner2; Hiltunen, J. Kalervo3;|
1Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, Finland
2Theodor Boveri Institute of Biosciences (Biocenter), University of Wurzburg, Germany
3Faculty of Biochemistry and Molecular Medicine, University of Oulu, Finland
4Biocenter Oulu, University of Oulu, Finland
|Online Access:||PDF Full Text (PDF, 2.7 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe2020120899853
International Union of Crystallography,
|Publish Date:|| 2020-12-08
The peroxisomal multifunctional enzyme type 1 (MFE1) catalyzes two successive reactions in the β-oxidation cycle: the 2E-enoyl-CoA hydratase (ECH) and NAD⁺-dependent 3S-hydroxyacyl-CoA dehydrogenase (HAD) reactions. MFE1 is a monomeric enzyme that has five domains. The N-terminal part (domains A and B) adopts the crotonase fold and the C-terminal part (domains C, D and E) adopts the HAD fold. A new crystal form of MFE1 has captured a conformation in which both active sites are noncompetent. This structure, at 1.7 Å resolution, shows the importance of the interactions between Phe272 in domain B (the linker helix; helix H10 of the crotonase fold) and the beginning of loop 2 (of the crotonase fold) in stabilizing the competent ECH active-site geometry. In addition, protein crystallographic binding studies using optimized crystal-treatment protocols have captured a structure with both the 3-ketodecanoyl-CoA product and NAD⁺ bound in the HAD active site, showing the interactions between 3-ketodecanoyl-CoA and residues of the C, D and E domains. Structural comparisons show the importance of domain movements, in particular of the C domain with respect to the D/E domains and of the A domain with respect to the HAD part. These comparisons suggest that the N-terminal part of the linker helix, which interacts tightly with domains A and E, functions as a hinge region for movement of the A domain with respect to the HAD part.
Acta Crystallographica. Section D, Structural biology
|Pages:||1256 - 1269|
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
1182 Biochemistry, cell and molecular biology
This work was supported by Instruct-ULTRA (Grant 731005), an EU H2020 project to further develop the services of Instruct-ERIC.
|EU Grant Number:||
(731005) INSTRUCT-ULTRA - Releasing the full potential of Instruct to expand and consolidate infrastructure services for integrated structural life science research
© International Union of Crystallography. The final authenticated version is available online at https://doi.org/10.1107/s2059798320013819.