Title Structural and sequence comparisons of bacterial enoyl-CoA isomerase and enoyl-CoA hydratase
Author Jisub Hwang1,2†, Chang-Sook Jeong1,2†, Chang Woo Lee1†, Seung Chul Shin3, Han-Woo Kim1,2, Sung Gu Lee1,2, Ui Joung Youn2,3, Chang Sup Lee4, Tae-Jin Oh5,6,7, Hak Jun Kim8, Hyun Park9*, Hyun Ho Park10*, and Jun Hyuck Lee1,2*
Address 1Unit of Research for Practical Application, Korea Polar Research Institute, Incheon 21990, Republic of Korea, 2Department of Polar Sciences, University of Science and Technology, Incheon 21990, Republic of Korea, 3Division of Life Sciences, Korea Polar Research Institute, Incheon 21990, Republic of Korea, 4College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea, 5Department of Life Science and Biochemical Engineering, Graduate School, SunMoon University, Asan 31460, Republic of Korea, 6Genome-based BioIT Convergence Institute, Asan 31460, Republic of Korea, 7Department of Pharmaceutical Engineering and Biotechnology, SunMoon University, Asan 31460, Republic of Korea, 8Department of Chemistry, Pukyong National University, Busan 48513, Republic of Korea, 9Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea, 10College of Pharmacy, Chung-Ang University, Seoul 06974, Republic of Korea
Bibliography Journal of Microbiology, 58(7),606–613, 2020,
DOI 10.1007/s12275-020-0089-1
Key Words crystal structure, enoyl-CoA isomerase, enoyl- CoA hydratase, X-ray crystallography
Abstract Crystal structures of enoyl-coenzyme A (CoA) isomerase from Bosea sp. PAMC 26642 (BoECI) and enoyl-CoA hydratase from Hymenobacter sp. PAMC 26628 (HyECH) were determined at 2.35 and 2.70 Å resolution, respectively. BoECI and HyECH are members of the crotonase superfamily and are enzymes known to be involved in fatty acid degradation. Structurally, these enzymes are highly similar except for the orientation of their C-terminal helix domain. Analytical ultracentrifugation was performed to determine the oligomerization states of BoECI and HyECH revealing they exist as trimers in solution. However, their putative ligand-binding sites and active site residue compositions are dissimilar. Comparative sequence and structural analysis revealed that the active site of BoECI had one glutamate residue (Glu135), this site is occupied by an aspartate in some ECIs, and the active sites of HyECH had two highly conserved glutamate residues (Glu118 and Glu138). Moreover, HyECH possesses a salt bridge interaction between Glu98 and Arg152 near the active site. This interaction may allow the catalytic Glu118 residue to have a specific conformation for the ECH enzyme reaction. This salt bridge interaction is highly conserved in known bacterial ECH structures and ECI enzymes do not have this type of interaction. Collectively, our comparative sequential and structural studies have provided useful information to distinguish and classify two similar bacterial crotonase superfamily enzymes.