Coenzyme binding in F420-dependent secondary alcohol dehydrogenase, a member of the bacterial luciferase family

• Published in: Structure (London) Vol. 12; no. 3; pp. 361 - 370
• Main Authors: Aufhammer, Stephan W, Warkentin, Eberhard, Berk, Holger, Shima, Seigo, Thauer, Rudolf K, Ermler, Ulrich
• Format: Journal Article
• Language: English
• Published: United States 01.03.2004
• Subjects: Acetone - metabolism; Alcohol Oxidoreductases - chemistry; Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - metabolism; Amino Acid Sequence; Binding Sites; Coenzymes - metabolism; Crystallography, X-Ray; Dimerization; Luciferases - genetics; Methanomicrobiaceae - enzymology; Methanomicrobiaceae - genetics; Methanomicrobiaceae - metabolism; Molecular Sequence Data; Multigene Family; Protein Binding; Protein Structure, Tertiary; Sequence Alignment
• ISSN: 0969-2126
• DOI: 10.1016/j.str.2004.02.010

F(420)-dependent secondary alcohol dehydrogenase (Adf) from methanogenic archaea is a member of the growing bacterial luciferase family which are all TIM barrel enzymes, most of which with an unusual nonprolyl cis peptide bond. We report here on the crystal structure of Adf from Methanoculleus thermophilicus at 1.8 A resolution in complex with a F(420)-acetone adduct. The knowledge of the F(420) binding mode in Adf provides the molecular basis for modeling F(420) and FMN into the other enzymes of the family. A nonprolyl cis peptide bond was identified as an essential part of a bulge that serves as backstop at the Re-face of F(420) to keep it in a bent conformation. The acetone moiety of the F(420)-acetone adduct is positioned at the Si-face of F(420) deeply buried inside the protein. Isopropanol can be reliably modeled and a hydrogen transfer mechanism postulated. His39 and Glu108 can be identified as key players for binding of the acetone or isopropanol oxygens and for catalysis.