The importance of the conserved Arg191–Asp227 salt bridge of triosephosphate isomerase for folding, stability, and catalysis

• Published in: FEBS letters 2002-05, Vol.518 (1), p.39-42
• Main Authors: Kursula, Inari, Partanen, Sanna, Lambeir, Anne-Marie, Wierenga, Rik K.
• Format: Article
• Language: English
• Subjects: Animals; Arginine - genetics; Arginine - physiology; Aspartic Acid - genetics; Aspartic Acid - physiology; Binding Sites; Catalysis; Conserved Sequence; DHAP, dihydroxyacetone phosphate; Enzyme Stability; Folding; GAP, glyceraldehyde-3-phosphate; GuHCl, guanidinium hydrochloride; Hydrogen Bonding; Kinetics; Models, Chemical; Models, Molecular; Mutation; PGA, 2-phosphoglycolate; Protein Folding; Salt bridge; Salts - chemistry; Stability; TIM, triosephosphate isomerase; Triose-Phosphate Isomerase - chemistry; Triose-Phosphate Isomerase - genetics; Triose-Phosphate Isomerase - metabolism; Triosephosphate isomerase; Trypanosoma brucei brucei - enzymology
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(02)02639-X

Triosephosphate isomerase (TIM) has a conserved salt bridge 20 Å away from both the active site and the dimer interface. In this study, four salt bridge mutants of Trypanosoma brucei brucei TIM were characterized. The folding and stability of the mutants are impaired compared to the wild-type enzyme. This salt bridge is part of a hydrogen bonding network which tethers the C-terminal β7α7β8α8 unit to the bulk of the protein. In the variants D227N, D227A, and R191S, this network is preserved, as can be deduced from the structure of the R191S variant. In the R191A variant, the side chain at position 191 cannot contribute to this network. Also the catalytic power of this variant is most affected.