A comparative study of the kinetics and stereochemistry of the serine hydroxymethyltransferase- and tryptophan synthase-catalysed exchange of the pro-2 R and pro-2 S protons of glycine

• Published in: Biochemical journal 1991-03, Vol.274 (3), p.807-812
• Main Authors: Malthouse, J P G, Milne, J J, Gariani, L S
• Format: Article
• Language: English
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/bj2740807

The stereospecificity of the serine hydroxymethyltransferase (EC 2.1.2.1)- and tryptophan synthase (EC 4.2.1.20)- catalysed exchange of the pro-2R and pro-2S alpha-protons of glycine was investigated by using 13C n.m.r. The exchange process is described in terms of a minimal four-step mechanism, and a method for analysing the exchange process by complete progress curves is presented. It is shown that serine hydroxymethyltransferase does not have absolute stereospecificity for the pro-2S-proton of glycine, but it catalyses the exchange of this proton 7400 times faster than the pro-2R proton of glycine. Tryptophan synthase is shown preferentially to catalyse the exchange of the pro-2R proton of glycine at a rate 380 times faster than the pro-2S proton of glycine. The exchange rates for the rapidly exchanged alpha-protons of glycine are similar for both enzymes. However, the exchange rates of the slowly exchanged alpha-protons differ by an order of magnitude. The structural features that may be responsible for the differences in the stereospecificity of the two enzymes are discussed.