Structure determination of glycogen synthase kinase-3 from Leishmania major and comparative inhibitor structure-activity relationships with Trypanosoma brucei GSK-3

Mol Biochem Parasitol. 2011 Apr;176(2):98-108. doi: 10.1016/j.molbiopara.2010.12.009. Epub 2010 Dec 30.

Abstract

Glycogen synthase kinase-3 (GSK-3) is a drug target under intense investigation in pharmaceutical companies and constitutes an attractive piggyback target for eukaryotic pathogens. Two different GSKs are found in trypanosomatids, one about 150 residues shorter than the other. GSK-3 short (GeneDB: Tb927.10.13780) has previously been validated genetically as a drug target in Trypanosoma brucei by RNAi induced growth retardation; and chemically by correlation between enzyme and in vitro growth inhibition. Here, we report investigation of the equivalent GSK-3 short enzymes of L. major (LmjF18.0270) and L. infantum (LinJ18_V3.0270, identical in amino acid sequences to LdonGSK-3 short) and a crystal structure of LmajGSK-3 short at 2 Å resolution. The inhibitor structure-activity relationships (SARs) of L. major and L. infantum are virtually identical, suggesting that inhibitors could be useful for both cutaneous and visceral leishmaniasis. Leishmania spp. GSK-3 short has different inhibitor SARs than TbruGSK-3 short, which can be explained mostly by two variant residues in the ATP-binding pocket. Indeed, mutating these residues in the ATP-binding site of LmajGSK-3 short to the TbruGSK-3 short equivalents results in a mutant LmajGSK-3 short enzyme with SAR more similar to that of TbruGSK-3 short. The differences between human GSK-3β (HsGSK-3β) and LmajGSK-3 short SAR suggest that compounds which selectively inhibit LmajGSK-3 short may be found.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Motifs / drug effects
  • Binding Sites
  • Crystallography, X-Ray
  • Escherichia coli
  • Gene Expression
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors*
  • Glycogen Synthase Kinase 3 / chemistry
  • Glycogen Synthase Kinase 3 / genetics
  • Glycogen Synthase Kinase 3 / metabolism
  • Humans
  • Kinetics
  • Leishmania infantum / drug effects
  • Leishmania infantum / genetics
  • Leishmania infantum / metabolism
  • Leishmania major / drug effects
  • Leishmania major / genetics
  • Leishmania major / metabolism
  • Leishmaniasis, Cutaneous / drug therapy
  • Leishmaniasis, Cutaneous / genetics
  • Leishmaniasis, Cutaneous / metabolism
  • Leishmaniasis, Visceral / drug therapy
  • Leishmaniasis, Visceral / genetics
  • Leishmaniasis, Visceral / metabolism
  • Models, Molecular
  • Mutation
  • Protein Binding / drug effects
  • Protein Isoforms / antagonists & inhibitors*
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Kinase Inhibitors / pharmacology*
  • Protein Kinase Inhibitors / therapeutic use
  • Recombinant Proteins / antagonists & inhibitors*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Species Specificity
  • Structure-Activity Relationship
  • Trypanosoma brucei brucei / drug effects
  • Trypanosoma brucei brucei / genetics
  • Trypanosoma brucei brucei / metabolism

Substances

  • Protein Isoforms
  • Protein Kinase Inhibitors
  • Recombinant Proteins
  • Glycogen Synthase Kinase 3