Dynamic origins of substrate promiscuity in bacterial galactokinases

Margaret McAuley, Meilan Huang, David Timson

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Galactokinase catalyses the ATP-dependent phosphorylation of galactose and structurally related sugars. The enzyme has attracted interest as a potential biocatalyst for the production of sugar 1-phosphates and several attempts have been made to broaden its specificity. In general, bacterial galactokinases have wider substrate ranges than mammalian ones. The enzymes from Escherichia coli and Lactococcus lactis have received particular attention and a number of variants with increased promiscuity have been identified. Here, we present a molecular dynamics study designed to investigate the molecular causes of the wider
substrate ranges of these enzymes and their variants with particular reference to protein mobility. Some regions close to the active site of the enzyme have different structures in the bacterial enzymes compared to the human one. Alterations known to increase the substrate range (e.g. Y371H in the E. coli enzyme),
tend to alter the conformation of a key α-helical region (residues 216-232 in the E. coli enzyme). The equivalent helix in the human enzyme has previously been predicted to be altered in variants which affect catalytic activity or protein stability. This helix appears to be a key region in galactokinases from a range of
species and may represent an interesting target for future attempts to broaden the specificity of galactokinases.
Original languageEnglish
Article number107839
JournalCarbohydrate Research
Volume486
DOIs
Publication statusPublished - 17 Oct 2019

Fingerprint

Enzymes
Escherichia coli
Galactokinase
Sugar Phosphates
Bacterial Structures
Lactococcus lactis
Protein Stability
Molecular Dynamics Simulation
Galactose
Catalytic Domain
Adenosine Triphosphate
Phosphorylation
Proteins

Keywords

  • GHMP kinase
  • Molecular dynamics
  • Biocatalyst
  • Protein mobility
  • Enzyme specificity
  • Sugar 1-phosphate

Cite this

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Dynamic origins of substrate promiscuity in bacterial galactokinases. / McAuley, Margaret; Huang, Meilan; Timson, David.

In: Carbohydrate Research, Vol. 486, 107839, 17.10.2019.

Research output: Contribution to journalArticleResearchpeer-review

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