title: A hydrogen-bonding network in mammalian sorbitol dehydrogenase stabilizes the tetrameric state and is essential for the catalytic power.
abstract: Subunit interaction in sorbitol dehydrogenase (SDH) has been studied with in vitro and in silico methods identifying a vital hydrogen-bonding network, which is strictly conserved among mammalian SDH proteins. Mutation of one of the residues in the hydrogen-bonding network, Tyr110Phe, abolished the enzymatic activity and destabilized the protein into tetramers, dimers and monomers as judged from gel filtration experiments at different temperatures compared to only tetramers for the wild-type protein below 307 K. The determined equilibrium constants revealed a large difference in Gibbs energy (8 kJ/mol) for the tetramer stability between wild-type SDH and the mutated form Tyr110Phe SDH. The results focus on a network of coupled hydrogen bonds in wild-type SDH that uphold the protein interface, which is specific and favorable to electrostatic, van der Waals and hydrogen-bond interactions between subunits, interactions that are crucial for the catalytic power of SDH.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden.
authors.forename: M
authors.initials: M
authors.lastname: Hellgren
authors.forename: C
authors.initials: C
authors.lastname: Kaiser
authors.forename: S
authors.initials: S
authors.lastname: de Haij
authors.forename: A
authors.initials: A
authors.lastname: Norberg
authors.forename: J-O
authors.initials: JO
authors.lastname: Höög
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D004735:Energy Transfer; D004795:Enzyme Stability; D006860:Hydrogen Bonding; D066298:In Vitro Techniques; D007064:L-Iditol 2-Dehydrogenase / Q000737:chemistry*; D008956:Models, Chemical; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins; D017386:Sequence Homology, Amino Acid
chemical_list: D011994:Recombinant Proteins; D007064:L-Iditol 2-Dehydrogenase