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

11 Citations (Scopus)

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

Fingerprint

Muscle
Muscles
Stabilization
Synthetic Genes
Kinetics
Enzymes
Glutamine
Escherichia coli
Nuclear magnetic resonance spectroscopy
Tyrosine
Arginine
Conformations
Structural properties
Catalytic Domain
Bone
Magnetic Resonance Spectroscopy
Genes
Phosphates
Bone and Bones
acylphosphatase

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

Cite this

Taddei, N., Modesti, A., Bucciantini, M., Stefani, M., Magherini, F., Vecchi, M., ... 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

Properties of N-terminus truncated and C-terminus mutated muscle acylphosphatases. / Taddei, Niccolò; Modesti, Alessandra; Bucciantini, Monica; Stefani, Massimo; Magherini, Francesca; Vecchi, Manuela; Raugei, Giovanni; Ramponi, Giampietro.

In: FEBS Letters, Vol. 362, No. 2, 03.04.1995, p. 175-179.

Research output: Contribution to journalArticle

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, vol. 362, no. 2, pp. 175-179. https://doi.org/10.1016/0014-5793(95)00236-3
Taddei, Niccolò ; Modesti, Alessandra ; Bucciantini, Monica ; Stefani, Massimo ; Magherini, Francesca ; Vecchi, Manuela ; Raugei, Giovanni ; Ramponi, Giampietro. / Properties of N-terminus truncated and C-terminus mutated muscle acylphosphatases. In: FEBS Letters. 1995 ; Vol. 362, No. 2. pp. 175-179.
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