title: Emergence of pyridoxal phosphorylation through a promiscuous ancestor during the evolution of hydroxymethyl pyrimidine kinases.
abstract: In the family of ATP-dependent vitamin kinases, several bifunctional enzymes that phosphorylate hydroxymethyl pyrimidine (HMP) and pyridoxal (PL) have been described besides enzymes specific towards HMP. To determine how bifunctionality emerged, we reconstructed the sequence of three ancestors of HMP kinases, experimentally resurrected, and assayed the enzymatic activity of their last common ancestor. The latter has ∼ 8-fold higher specificity for HMP due to a glutamine residue (Gln44) that is a key determinant of the specificity towards HMP, although it is capable of phosphorylating both substrates. These results show how a specific enzyme with catalytic promiscuity gave rise to current bifunctional enzymes.
authors: Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
authors: Victor
authors: V
authors: Castro-Fernandez
authors: Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
authors: Felipe
authors: F
authors: Bravo-Moraga
authors: Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
authors: Cesar A
authors: CA
authors: Ramirez-Sarmiento
authors: Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile. Electronic address: vguixe@uchile.cl.
authors: Victoria
authors: V
authors: Guixe
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D055162:Biocatalysis; D019143:Evolution, Molecular; D006801:Humans; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000378:metabolism*; D010802:Phylogeny; D011487:Protein Conformation; D011730:Pyridoxal / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D011730:Pyridoxal; D000255:Adenosine Triphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C059599:hydroxymethylpyrimidine kinase
keywords: Bifunctional enzyme; Enzyme evolution; HMP kinase; Pyridoxal kinase; Resurrection ancestral enzyme