Structure/function relationship studies on the T/S residues 173-177 of rat ODC

José Manuel Matés, Alicia E. Del Valle, José Luis Urdiales, Catherine Coleman, David Feith, M. Teresa Olmo, Anthony Pegg, Francisca Sánchez-Jiménez

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Abstract

A well-conserved T/S cluster was detected among vertebrate ornithine decarboxylase by computer analysis (E. Viguera, O. Trelles, J.L. Urdiales, J.M. Mates, F. Sanchez-Jimenez, Trends Biochem. Sci. 19 (1994) 318-319). In the present report we studied the role of these residues (173, 176 and 177 in rat ornithine decarboxylase (ODC)) in enzymic activity and stability by in vitro expression, kinetic characterization and in vitro degradation of site-directed mutants. These T/S residues are substituted by a D/E-enriched fragment in other lower eukaryotic ODCs. The substitution of the T/S-enriched fragment (TLKTS) of rat ODC by the negative charged fragment of T. brucei ODC (KVEDC) did not affect protein stability, but increased K(m) values of the mutant enzyme. The substitution of the T/S residues by alanine also has a similar effect on rat ODC kinetic values. However, results indicate that polarity of the fragment must be an important factor for protein conformation, since the latter mutant, having no T/S or D/E residue in the fragment (ALKAA), showed reduced stability in vitro. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)113-120
Number of pages8
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume1386
Issue number1
DOIs
StatePublished - Jul 28 1998

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology

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    Matés, J. M., Del Valle, A. E., Urdiales, J. L., Coleman, C., Feith, D., Olmo, M. T., Pegg, A., & Sánchez-Jiménez, F. (1998). Structure/function relationship studies on the T/S residues 173-177 of rat ODC. Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, 1386(1), 113-120. https://doi.org/10.1016/S0167-4838(98)00090-9