Remote interactions explain the unusual regioselectivity of lipase from Pseudomonas cepacia toward the secondary hydroxyl of 2′-deoxynucleosides

Iván Lavandera, Susana Fernández, Julia Magdalena, Miguel Ferrero, Harjap Grewal, Christopher K. Savile, Romas J. Kazlauskas, Vicente Gotor

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

Lipase from Pseudomonas cepacia (PCL) surprisingly favors acylation of the secondary hydroxyl at the 3′-position over the primary hydroxyl at the 5′-position in 2′-deoxynucleosides by up to >98:1. Catalytically productive tetrahedral intermediate analogues for both orientations were found by molecular modeling. However, acylation of the 3′-hydroxyl places the thymine base in the alternate hydrophobic pocket of PCL's substrate-binding site where it can hydrogen bond to the side-chain hydroxyls of Tyr23 and Tyr29 and the main chain carbonyl of Leu17. Conversely, acylation of the 5′-hydroxyl leaves the thymine base in the solvent where there is no favorable binding to the enzyme. We propose that these remote stabilizing interactions between the thymine base and PCL's substrate-binding site stabilize the 3′-acylation transition state and thus account for the unusual regioselectivity.

Original languageEnglish (US)
Pages (from-to)693-698
Number of pages6
JournalChemBioChem
Volume7
Issue number4
DOIs
StatePublished - Apr 2006

Keywords

  • Biotransformations
  • Enzyme catalysis
  • Molecular modeling
  • Nucleosides
  • Regioselective acylations

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