From amino acid to glucosinolate biosynthesis: protein sequence changes in the evolution of methylthioalkylmalate synthase in Arabidopsis

Plant Cell. 2011 Jan;23(1):38-53. doi: 10.1105/tpc.110.079269. Epub 2011 Jan 4.


Methylthioalkylmalate synthase (MAM) catalyzes the committed step in the side chain elongation of Met, yielding important precursors for glucosinolate biosynthesis in Arabidopsis thaliana and other Brassicaceae species. MAM is believed to have evolved from isopropylmalate synthase (IPMS), an enzyme involved in Leu biosynthesis, based on phylogenetic analyses and an overlap of catalytic abilities. Here, we investigated the changes in protein structure that have occurred during the recruitment of IPMS from amino acid to glucosinolate metabolism. The major sequence difference between IPMS and MAM is the absence of 120 amino acids at the C-terminal end of MAM that constitute a regulatory domain for Leu-mediated feedback inhibition. Truncation of this domain in Arabidopsis IPMS2 results in loss of Leu feedback inhibition and quaternary structure, two features common to MAM enzymes, plus an 8.4-fold increase in the k(cat)/K(m) for a MAM substrate. Additional exchange of two amino acids in the active site resulted in a MAM-like enzyme that had little residual IPMS activity. Hence, combination of the loss of the regulatory domain and a few additional amino acid exchanges can explain the evolution of MAM from IPMS during its recruitment from primary to secondary metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 2-Isopropylmalate Synthase / genetics
  • 2-Isopropylmalate Synthase / metabolism
  • Amino Acid Sequence
  • Arabidopsis / enzymology
  • Arabidopsis / genetics*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Catalytic Domain
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Plant
  • Glucosinolates / biosynthesis*
  • Molecular Sequence Data
  • Oxo-Acid-Lyases / genetics
  • Oxo-Acid-Lyases / metabolism*
  • Protein Structure, Quaternary
  • Sequence Alignment
  • Sequence Analysis, Protein
  • Substrate Specificity


  • Arabidopsis Proteins
  • Glucosinolates
  • 2-Isopropylmalate Synthase
  • Oxo-Acid-Lyases
  • methylthioalkylmalate synthase 1