title: Mechanism for folate-independent aldolase reaction catalyzed by serine hydroxymethyltransferase.
abstract: Serine hydroxymethyltransferase catalyzes the cleavage of β-hydroxyamino acids into glycine and aldehydes in the absence of tetrahydrofolate. The enzyme accepts various β-hydroxyamino acids as the substrate of this reaction. The reaction rate varies depending on the substituent and stereochemistry at the Cβ atom: the erythro forms and the β-phenyl substituent are preferred over the threo forms and the β-methyl substituent, respectively. Although several mechanisms have been proposed, what determines the substrate preference remains unclear. We first performed quantum mechanical calculations to assess the validity of the reaction mechanisms. The results indicate that the retro-aldol mechanism starting with abstraction of the proton from the β-hydroxyl group is plausible. This also suggests that Cα-Cβ bond cleavage is the rate-limiting step. We next measured the dependence of the rate constants on temperature with four representative substrates and calculated the activation energies and pre-exponential factors from the Arrhenius plots. The activation energies of the erythro forms were lower than those of the threo forms. The β-phenyl substituent lowered the activation energy in the threo form, whereas it did not alter the activation energy but increased the pre-exponential factor in the erythro form. We present a unified model to explain the origin of the substituent and stereochemical preferences by combining the theoretical and experimental results. A possible biological role of the tetrahydrofolate-independent activity in thermophiles is also discussed.
authors.affiliation: Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Chiba
authors.forename: Tohru
authors.initials: T
authors.lastname: Terada
authors.forename: Masafumi
authors.initials: M
authors.lastname: Kameya
authors.forename: Kentaro
authors.initials: K
authors.lastname: Shimizu
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Arai
authors.forename: Masaharu
authors.initials: M
authors.lastname: Ishii
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Igarashi
mesh_terms: D001419:Bacteria / Q000201:enzymology*; D055162:Biocatalysis; D005492:Folic Acid; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D012696:Glycine Hydroxymethyltransferase / Q000378:metabolism*; D007700:Kinetics; D008956:Models, Chemical; D008958:Models, Molecular; D013379:Substrate Specificity; D013696:Temperature; D013763:Tetrahydrofolates; D013816:Thermodynamics
chemical_list: D013763:Tetrahydrofolates; C030371:5,6,7,8-tetrahydrofolic acid; D005492:Folic Acid; D012696:Glycine Hydroxymethyltransferase; D005634:Fructose-Bisphosphate Aldolase