An engineered variant of MECR reductase reveals indispensability of long-chain acyl-ACPs for mitochondrial respiration

DOI: 10.1038/s41467-023-36358-7

Authors: M. Tanvir Rahman (University of Oulu) , M. Kristian Koski (University of Oulu) , Joanna Panecka-Hofman (University of Warsaw; Heidelberg Institute for Theoretical Studies (HITS)) , Werner Schmitz (University of Würzburg) , Alexander J. Kastaniotis (University of Oulu) , Rebecca C. Wade (Heidelberg Institute for Theoretical Studies (HITS); Heidelberg University) , R. K. Wierenga (University of Oulu) , J. Kalervo Hiltunen (University of Oulu) , Kaija J. Autio (University of Oulu)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 14

State: Published (Approved)
Published: February 2023
Diamond Proposal Number(s): 23346

Abstract: Mitochondrial fatty acid synthesis (mtFAS) is essential for respiratory function. MtFAS generates the octanoic acid precursor for lipoic acid synthesis, but the role of longer fatty acid products has remained unclear. The structurally well-characterized component of mtFAS, human 2E-enoyl-ACP reductase (MECR) rescues respiratory growth and lipoylation defects of a Saccharomyces cerevisiae Δetr1 strain lacking native mtFAS enoyl reductase. To address the role of longer products of mtFAS, we employed in silico molecular simulations to design a MECR variant with a shortened substrate binding cavity. Our in vitro and in vivo analyses indicate that the MECR G165Q variant allows synthesis of octanoyl groups but not long chain fatty acids, confirming the validity of our computational approach to engineer substrate length specificity. Furthermore, our data imply that restoring lipoylation in mtFAS deficient yeast strains is not sufficient to support respiration and that long chain acyl-ACPs generated by mtFAS are required for mitochondrial function.

Journal Keywords: Enzyme mechanisms; Fatty acids; Protein design; Respiration; X-ray crystallography

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials


Instruments: I04-Macromolecular Crystallography

Added On: 08/02/2023 09:48

Documents:
s41467-023-36358-7.pdf

Discipline Tags:

Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)