title: Gly387 of murine ornithine decarboxylase is essential for the formation of stable homodimers.
abstract: In its active form mammalian ornithine decarboxylase (ODC) is a homodimer composed of two 53-kDa subunits while the monomer retains no enzymic activity. In the present study we demonstrate that Gly387 of mouse ODC plays an important role in enabling dimer formation. Gly387 of mouse ODC, an evolutionary conserved residue, was converted to all possible 19 amino acids using site-directed mutagenesis. With the exception of alanine, all other substitutions of Gly387 completely abolished enzymic activity. Cross-linking analysis and fractionation through a Superose-12 sizing column have demonstrated that mutant subunits are detected only in their monomeric form. These results strongly suggest that the primary lesion of substitution at position 387 of mouse ODC is the inability of mutant subunits to associate with each other to form the active homodimers. In agreement with this conclusion, G387A, the only mutant that retained partial activity, displayed reduced dimerization. The degradation rate of ODC mutants in which Gly387 was substituted by aspartic acid or alanine was enhanced compared to the wild-type enzyme, suggesting that monomers may be more susceptible to degradation.
authors.affiliation: Department of Molecular Genetics and Virology, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: K E
authors.initials: KE
authors.lastname: Tobias
authors.forename: E
authors.initials: E
authors.lastname: Mamroud-Kidron
authors.forename: C
authors.initials: C
authors.lastname: Kahana
mesh_terms: D000409:Alanine / Q000737:chemistry; D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D005998:Glycine / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009838:Oligodeoxyribonucleotides; D009955:Ornithine Decarboxylase / Q000737:chemistry* / Q000378:metabolism
chemical_list: D009838:Oligodeoxyribonucleotides; D001224:Aspartic Acid; D009955:Ornithine Decarboxylase; D000409:Alanine; D005998:Glycine