Monomeric S-adenosylmethionine decarboxylase from plants provides an alternative to putrescine stimulation

Biochemistry. 2002 Dec 10;41(49):14509-17. doi: 10.1021/bi026710u.

Abstract

S-Adenosylmethionine decarboxylase has been implicated in cell growth and differentiation and is synthesized as a proenzyme, which undergoes autocatalytic cleavage to generate an active site pyruvoyl group. In mammals, S-adenosylmethionine decarboxylase is active as a dimer in which each protomer contains one alpha subunit and one beta subunit. In many higher organisms, autocatalysis and decarboxylation are stimulated by putrescine, which binds in a buried site containing numerous negatively charged residues. In contrast, plant S-adenosylmethionine decarboxylases are fully active in the absence of putrescine, with rapid autocatalysis that is not stimulated by putrescine. We have determined the structure of the S-adenosylmethionine decarboxylase from potato, Solanum tuberosum, to 2.3 A resolution. Unlike the previously determined human enzyme structure, the potato enzyme is a monomer in the crystal structure. Ultracentrifugation studies show that the potato enzyme is also a monomer under physiological conditions, with a weak self-association constant of 6.5 x 10(4) M(-)(1) for the monomer-dimer association. Although the potato enzyme contains most of the buried charged residues that make up the putrescine binding site in the human enzyme, there is no evidence for a putrescine binding site in the potato enzyme. Instead, several amino acid substitutions, including Leu13/Arg18, Phe111/Arg114, Asp174/Val181, and Phe285/His294 (human/potato), provide side chains that mimic the role of putrescine in the human enzyme. In the potato enzyme, the positively charged residues form an extensive network of hydrogen bonds bridging a cluster of highly conserved negatively charged residues and the active site, including interactions with the catalytic residues Glu16 and His249. The results explain the constitutively high activity of plant S-adenosylmethionine decarboxylases in the absence of putrescine and are consistent with previously proposed models for how putrescine together with the buried, negatively charged site regulates enzyme activity.

Publication types

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

MeSH terms

  • Adenosylmethionine Decarboxylase / chemistry*
  • Amino Acid Substitution
  • Animals
  • Binding Sites
  • Crystallography, X-Ray
  • Dimerization
  • Humans
  • Hydrogen Bonding
  • Models, Molecular
  • Plant Proteins / chemistry*
  • Protein Conformation
  • Protozoan Proteins / chemistry
  • Putrescine / chemistry*
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Solanum tuberosum / enzymology*
  • Static Electricity

Substances

  • Plant Proteins
  • Protozoan Proteins
  • Adenosylmethionine Decarboxylase
  • Putrescine

Associated data

  • PDB/1MHM