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

Sridhar, ShruthiFaculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland;Biocenter Oulu, University of Oulu, Oulu, Finland (author)

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

The peroxisomal multifunctional enzyme type 1 (MFE1) catalyzes two successive reactions in the beta-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 angstrom 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.

MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine hsv//eng
NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
peroxisomal multifunctional enzyme type 1
Rossmann fold
reaction mechanism
closed active site
fatty-acid oxidation
conformational flexibility.
Biology with specialization in Structural Biology
Biologi med inriktning mot strukturbiologi

Schmitz, Werner (author)
Hiltunen, J. Kalervo (author)
Venkatesan, Rajaram (author)
Bergmann, Ulrich (author)
Kiema, Tiila-Riikka (author)
Wierenga, Rikkert K. (author)
Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland;Biocenter Oulu, University of Oulu, Oulu, Finland (creator_code:org_t)

In:Acta Crystallographica Section D: International Union of Crystallography (IUCr)76:12, s. 1256-12692059-7983

By the author/editor: Sridhar, Shruthi; Schmitz, Werner; Hiltunen, J. Kal ...; Venkatesan, Raja ...; Bergmann, Ulrich; Kiema, Tiila-Rii ...; show more...; Wierenga, Rikker ...; show less...

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

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