title: Activity of α-Aminoadipate Reductase Depends on the N-Terminally Extending Domain.
abstract: L-α-Aminoadipic acid reductases catalyze the ATP- and NADPH-dependent reduction of L-α-aminoadipic acid to the corresponding 6-semialdehyde during fungal L-lysine biosynthesis. These reductases resemble peptide synthetases with regard to their multidomain composition but feature a unique domain of elusive function--now referred to as an adenylation activating (ADA) domain--that extends the reductase N-terminally. Truncated enzymes based on NPS3, the L-α-aminoadipic acid reductase of the basidiomycete Ceriporiopsis subvermispora, lacking the ADA domain either partially or entirely were tested for activity in vitro, together with an ADA-adenylation didomain and the ADA domainless adenylation domain. We provide evidence that the ADA domain is required for substrate adenylation: that is, the initial step of the catalytic turnover. Our biochemical data are supported by in silico modeling that identified the ADA domain as a partial peptide synthetase condensation domain.
authors.affiliation: Department of Pharmaceutical Microbiology, Hans-Knöll-Institut Friedrich-Schiller-Universität, Beutenbergstrasse 11a, 07745 Jena (Germany).
authors.forename: Daniel
authors.initials: D
authors.lastname: Kalb
authors.affiliation: Institute of Microbiology, Swiss Federal Institute of Technology (ETH), Vladimir-Prelog-Weg 1-5, 8093 Zürich (Switzerland).
authors.forename: Gerald
authors.initials: G
authors.lastname: Lackner
authors.affiliation: Department of Pharmaceutical Microbiology, Hans-Knöll-Institut Friedrich-Schiller-Universität, Beutenbergstrasse 11a, 07745 Jena (Germany).
authors.forename: Marcus
authors.initials: M
authors.lastname: Rappe
authors.affiliation: Department of Pharmaceutical Microbiology, Hans-Knöll-Institut Friedrich-Schiller-Universität, Beutenbergstrasse 11a, 07745 Jena (Germany). dirk.hoffmeister@hki-jena.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Hoffmeister
mesh_terms: D015074:2-Aminoadipic Acid / Q000378:metabolism; D001487:Basidiomycota / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D050647:L-Aminoadipate-Semialdehyde Dehydrogenase / Q000737:chemistry* / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism*; D008958:Models, Molecular; D017434:Protein Structure, Tertiary
chemical_list: D015074:2-Aminoadipic Acid; D050647:L-Aminoadipate-Semialdehyde Dehydrogenase; D008239:Lysine
keywords: amino acids; aminoadipate reductases; biosynthesis; lysine; nonribosomal peptide synthetases