NMR Studies of Solvent-Assisted Proton Transfer in a Biologically Relevant Schiff Base:  Toward a Distinction of Geometric and Equilibrium H-Bond Isotope Effects

• Published in: Journal of the American Chemical Society 2006-03, Vol.128 (10), p.3375-3387
• Main Authors: Sharif, Shasad, Denisov, Gleb S, Toney, Michael D, Limbach, Hans-Heinrich
• Format: Article
• Language: English
• Subjects: Aspartate Aminotransferases - chemistry; Biomimetic Materials - chemistry; Hydrogen Bonding; Kinetics; Methylamines - chemistry; Nuclear Magnetic Resonance, Biomolecular - methods; Pyridoxal Phosphate - chemistry; Schiff Bases - chemistry
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja056251v

The tautomeric equilibrium in a Schiff base, N-(3,5-dibromosalicylidene)-methylamine 1, a model for the hydrogen bonded structure of the cofactor pyridoxal-5‘-phosphate PLP which is located in the active site of the enzyme, was measured by means of 1H and 15N NMR and deuterium isotope effects on 15N chemical shifts at variable temperature and in different organic solvents. The position of the equilibrium was estimated using the one-bond 1 J(OHN) and vicinal 3 J(H αCNH) scalar coupling constants. Additionally, DFT calculations of a series of Schiff bases, N-(R1-salicylidene)-alkyl(R2)amines, were performed to obtain the hydrogen bond geometries. The latter made it possible to investigate a broad range of equilibrium positions. The increase of the polarity of the aprotic solvent shifts the proton in the intramolecular OHN hydrogen bond closer to the nitrogen. The addition of methanol and of hexafluoro-2-propanol to 1 in aprotic solvents models the PLP−water interaction in the enzymatic active site. The alcohols, which vary in acidity and change the polarity around the hydrogen bond, also stabilize the equilibrium, so that the proton is shifted to the nitrogen.