Emergence of pyridoxal phosphorylation through a promiscuous ancestor during the evolution of hydroxymethyl pyrimidine kinases. Issue 17 (19th June 2014)

• Record Type: Journal Article
• Authors: Castro-Fernandez, Victor; Bravo-Moraga, Felipe; Ramirez-Sarmiento, Cesar A.; Guixe, Victoria
• Is Part Of: FEBS letters. Volume 588:Issue 17(2014)
• Page Start: 3068
• Page End: 3073
• Publication Date: 2014-06-19
• Subjects: HMP -- 4-amino-5-hydroxymethyl-2-methylpyrimidine -- HMPK -- 4-amino-5-hydroxymethyl-2-methylpyrimidine kinase -- PL -- pyridoxal -- PLK -- pyridoxal kinase -- PLP -- pyridoxal-5′-phosphate -- THZK -- 4-methyl-5-β-hydroxy-ethylthiazole kinase -- HMP kinase -- Pyridoxal kinase -- Bifunctional enzyme -- Enzyme evolution -- Resurrection ancestral enzyme; Biochemistry -- Periodicals; Biophysics -- Periodicals; Molecular biology -- Periodicals; Biochimie -- Périodiques; Biochemistry; Biophysics; Molecular biology; Periodicals; 572.05
• DOI: 10.1016/j.febslet.2014.06.033
• Languages: English
• ISSNs: 0014-5793

Abstract

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. Abstract : Bifunctional HMPK/PLK enzymes are phylogenetically related to specific HMPK. Resurrection and experimental characterization of the last common ancestor of HMPKs. Last common ancestor of HMPKs was specific for HMP but capable of phosphorylating pyridoxal. PLK activity appeared in the HMPK group as a convergent event with respect to PLKs.