Quantitative functional characterization of conserved molecular interactions in the active site of mannitol 2-dehydrogenase

James E. Lucas, Justin Siegel

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Enzyme active site residues are often highly conserved, indicating a significant role in function. In this study we quantitate the functional contribution for all conserved molecular interactions occurring within a Michaelis complex for mannitol 2-dehydrogenase derived from Pseudomonas fluorescens (pfMDH). Through systematic mutagenesis of active site residues, we reveal that the molecular interactions in pfMDH mediated by highly conserved residues not directly involved in reaction chemistry can be as important to catalysis as those directly involved in the reaction chemistry. This quantitative analysis of the molecular interactions within the pfMDH active site provides direct insight into the functional role of each molecular interaction, several of which were unexpected based on canonical sequence conservation and structural analyses.

Original languageEnglish (US)
Pages (from-to)936-945
Number of pages10
JournalProtein Science
Volume24
Issue number6
DOIs
StatePublished - Jun 1 2015

Keywords

  • biing
  • catalysis
  • conserved
  • enzyme
  • interactions
  • mannitol 2-dehydrogenase
  • molecular
  • mutagenesis
  • quantitative
  • specificity

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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