Structural and functional analysis of a dimeric fumarylacetoacetate hydrolase (EaFAH) from psychrophilic Exiguobacterium antarcticum

Wanki Yoo; Chang Woo Lee; Boo-Young Kim; Ly Thi Huong Luu Le; Sun-Ha Park; Han-Woo Kim; Seung Chul Shin; Kyeong Kyu Kim; Jun Hyuck Lee; T Doohun Kim

doi:10.1016/j.bbrc.2018.12.183

Structural and functional analysis of a dimeric fumarylacetoacetate hydrolase (EaFAH) from psychrophilic Exiguobacterium antarcticum

Biochem Biophys Res Commun. 2019 Feb 12;509(3):773-778. doi: 10.1016/j.bbrc.2018.12.183. Epub 2019 Jan 8.

Authors

Affiliations

¹ Department of Chemistry, College of Natural Science, Sookmyung Women's University, Seoul, 04310, South Korea; Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon, 440-746, South Korea.
² Unit of Polar Genomics, Korea Polar Research Institute (KOPRI), Incheon, 21990, South Korea; Department of Polar Sciences, University of Science and Technology (UST), Incheon, 21990, South Korea.
³ Department of Chemistry, College of Natural Science, Sookmyung Women's University, Seoul, 04310, South Korea.
⁴ Unit of Polar Genomics, Korea Polar Research Institute (KOPRI), Incheon, 21990, South Korea.
⁵ Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon, 440-746, South Korea.
⁶ Unit of Polar Genomics, Korea Polar Research Institute (KOPRI), Incheon, 21990, South Korea; Department of Polar Sciences, University of Science and Technology (UST), Incheon, 21990, South Korea. Electronic address: junhyucklee@kopri.re.kr.
⁷ Department of Chemistry, College of Natural Science, Sookmyung Women's University, Seoul, 04310, South Korea. Electronic address: doohunkim@sookmyung.ac.kr.

PMID: 30630595
DOI: 10.1016/j.bbrc.2018.12.183

Abstract

Fumarylacetoacetate hydrolase (FAH) is essential for the degradation of aromatic amino acids as well as for the cleavage of carbon-carbon bonds in metabolites or small organic compounds. Here, the X-ray crystal structure of EaFAH, a dimeric fumarylacetoacetate hydrolase from Exiguobacterium antarcticum, was determined, and its functional properties were investigated using biochemical methods. EaFAH adopts a mixed β-sandwich roll fold with a highly flexible lid region (Val⁷³-Leu⁹⁴), and an Mg²⁺ ion is bound at the active site by coordinating to the three carboxylate oxygen atoms of Glu¹²⁴, Glu¹²⁶, and Asp¹⁵⁵. The hydrolytic activity of EaFAH toward various substrates, including linalyl acetate was investigated using native polyacrylamide gel electrophoresis, activity staining, gel filtration, circular dichroism spectroscopy, fluorescence, and enzyme assays.

Keywords: EaFAH; Exiguobacterium antarcticum; Fumarylacetoacetate hydrolase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Bacillaceae / chemistry*
Bacillaceae / genetics
Bacillaceae / metabolism
Bacterial Proteins / chemistry*
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Catalytic Domain
Crystallography, X-Ray
Hydrolases / chemistry*
Hydrolases / genetics
Hydrolases / metabolism
Hydrolysis
Magnesium / metabolism
Models, Molecular
Phylogeny
Protein Conformation
Protein Multimerization
Sequence Alignment
Substrate Specificity

Substances

Bacterial Proteins
Hydrolases
fumarylacetoacetase
Magnesium