Properties of N-terminus truncated and C-terminus mutated muscle acylphosphatases

Niccolò Taddei, Alessandra Modesti, Monica Bucciantini, Massimo Stefani, Francesca Magherini, Manuela Vecchi, Giovanni Raugei, Giampietro Ramponi

Research output: Contribution to journalArticle

Abstract

Enzymatic activity and structure of N-terminus truncated and C-terminus substituted muscle acylphosphatase mutants were investigated by kinetic studies under different conditions and 1H NMR spectroscopy, respectively. The N-terminus truncated mutant lacked the first six residues (Δ6), whereas arginine 97 and tyrosine 98 were replaced by glutamine giving two C-terminus substituted mutants (R97Q and Y98Q, respectively). All acylphosphatase forms were obtained by modifications of a synthetic gene coding for the human muscle enzyme which was expressed in E. coli. The Δ6 deletion mutant elicited a reduced specific activity and a native-like structure. The kinetic and structural properties of R97Q and Y98Q mutants indicate a possible role of Arg-97 in the stabilisation of the active site correct conformation, most likely via back-bone and side chain interactions with Arg-23, the residue involved in phosphate binding by the enzyme. This study also suggests a possible involvement of Tyr-98 in the stabilisation of the acylphosphatase overall structure.

Original languageEnglish
Pages (from-to)175-179
Number of pages5
JournalFEBS Letters
Volume362
Issue number2
DOIs
Publication statusPublished - Apr 3 1995

Keywords

  • Acylphosphatase H NMR spectrum
  • Acylphosphatase deletion mutant
  • Acylphosphatase mutant
  • Acylphosphatase recombinant

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Structural Biology

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    Taddei, N., Modesti, A., Bucciantini, M., Stefani, M., Magherini, F., Vecchi, M., Raugei, G., & Ramponi, G. (1995). Properties of N-terminus truncated and C-terminus mutated muscle acylphosphatases. FEBS Letters, 362(2), 175-179. https://doi.org/10.1016/0014-5793(95)00236-3