title: NADP-Dependent Aldehyde Dehydrogenase from ArchaeonPyrobaculum sp.1860: Structural and Functional Features.
abstract: We present the functional and structural characterization of the first archaeal thermostable NADP-dependent aldehyde dehydrogenase AlDHPyr1147.In vitro, AlDHPyr1147 catalyzes the irreversible oxidation of short aliphatic aldehydes at 60-85°С, and the affinity of AlDHPyr1147 to the NADP+ at 60°С is comparable to that for mesophilic analogues at 25°С. We determined the structures of the apo form of AlDHPyr1147 (3.04 Å resolution), three binary complexes with the coenzyme (1.90, 2.06, and 2.19 Å), and the ternary complex with the coenzyme and isobutyraldehyde as a substrate (2.66 Å). The nicotinamide moiety of the coenzyme is disordered in two binary complexes, while it is ordered in the ternary complex, as well as in the binary complex obtained after additional soaking with the substrate. AlDHPyr1147 structures demonstrate the strengthening of the dimeric contact (as compared with the analogues) and the concerted conformational flexibility of catalytic Cys287 and Glu253, as well as Leu254 and the nicotinamide moiety of the coenzyme. A comparison of the active sites of AlDHPyr1147 and dehydrogenases characterized earlier suggests that proton relay systems, which were previously proposed for dehydrogenases of this family, are blocked in AlDHPyr1147, and the proton release in the latter can occur through the substrate channel.
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia.
authors.forename: Ekaterina Yu
authors.identifier: 0000-0002-4954-7547
authors.initials: EY
authors.lastname: Bezsudnova
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia; Institute of Mathematical Problems of Biology RAS, The Branch of Keldysh Institute of Applied Mathematics of the Russian Academy of Sciences, Pushchino, Russia.
authors.forename: Tatiana E
authors.initials: TE
authors.lastname: Petrova
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia.
authors.forename: Natalia V
authors.initials: NV
authors.lastname: Artemova
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia; NBICS Center, National Research Centre "Kurchatov Institute", Akad. Kurchatova Sqr. 1, Moscow 123182, Russia.
authors.forename: Konstantin M
authors.initials: KM
authors.lastname: Boyko
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia.
authors.forename: Ivan G
authors.initials: IG
authors.lastname: Shabalin
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia; NBICS Center, National Research Centre "Kurchatov Institute", Akad. Kurchatova Sqr. 1, Moscow 123182, Russia.
authors.forename: Tatiana V
authors.initials: TV
authors.lastname: Rakitina
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia; Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, Vavilov Str. 32, Moscow 119991, Russia.
authors.forename: Konstantin M
authors.initials: KM
authors.lastname: Polyakov
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, Russia; NBICS Center, National Research Centre "Kurchatov Institute", Akad. Kurchatova Sqr. 1, Moscow 123182, Russia.
authors.forename: Vladimir O
authors.initials: VO
authors.lastname: Popov
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000737:chemistry* / Q000378:metabolism*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004795:Enzyme Stability; D006358:Hot Temperature; D007700:Kinetics; D008958:Models, Molecular; D009249:NADP / Q000378:metabolism*; D011487:Protein Conformation; D041543:Pyrobaculum / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D009249:NADP; D000444:Aldehyde Dehydrogenase