Insights into acylphosphatase structure and catalytic mechanism

Stefani, Massimo; Taddei, Niccolò; Ramponi, Giampietro

doi:10.1007/pl00000585

Cited by 84 publications

(94 citation statements)

References 74 publications

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“…EcoAcP displays substrate and phosphate affinities typical of the AcP family (see Table 1), and a pH optimum in agreement with the proposed catalytic mechanism [1]. However, its k cat value is very low when compared to that of the other AcPs (except AcPDro) including hyperthermophilic AcPs assayed at room temperature.…”

Section: Discussionsupporting

confidence: 76%

The intrachain disulfide bridge is responsible of the unusual stability properties of novel acylphosphatase from Escherichia coli

Ramazzotti¹,

Parrini²,

Stefani³

et al. 2006

FEBS Letters

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Acylphosphatase (AcP) activity in prokaryotes was classically attributed to some aspecific acid phosphatases. We identified an open reading frame for a putative AcP in the b0968 Escherichia coli gene and purified the recombinant enzyme after checking by RT-PCR that it was indeed expressed. EcoAcP has a predicted typical fold of the AcP family but displays a very low specific activity and a high structural stability differently from its mesophilic and similarly to its hyperthermophilic counterparts. Site directed mutagenesis suggests that, together with other structural features, the intrachain S-S bridge in EcoAcP is involved in a remarkable thermal and chemical stabilization of the protein without affecting its catalytic activity.

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Section: Discussionsupporting

confidence: 76%

The intrachain disulfide bridge is responsible of the unusual stability properties of novel acylphosphatase from Escherichia coli

Ramazzotti¹,

Parrini²,

Stefani³

et al. 2006

FEBS Letters

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“…Acylphosphatase (AcP, EC 3.6.1.7), a widespread enzyme that is found from archaebacteria to human, specifically catalyses the hydrolysis of carboxyl-phosphate bond in acyl phosphates [1]. The true physiological function of AcP has not been fully revealed.…”

Section: Introductionmentioning

confidence: 99%

Solution structure and conformational heterogeneity of acylphosphatase from Bacillus subtilis

et al. 2010

FEBS Letters

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a b s t r a c tAcylphosphatase is a small enzyme that catalyzes the hydrolysis of acyl phosphates. Here, we present the solution structure of acylphosphatase from Bacillus subtilis (BsAcP), the first from a Gram-positive bacterium. We found that its active site is disordered, whereas it converted to an ordered state upon ligand binding. The structure of BsAcP is sensitive to pH and it has multiple conformations in equilibrium at acidic pH (pH < 5.8). Only one main conformation could bind ligand, and the relative population of these states is modulated by ligand concentration. This study provides direct evidence for the role of ligand in conformational selection.

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“…3 CT AcP is a singledomain protein of 98 residues and constitutes a simple model system for equilibrium and kinetic studies of folding. CT AcP catalyzes the hydrolysis of acylphosphates of physiological relevance such as 1,3-bisphosphoglycerate, carbamoylphosphate, and ␤-aspartylphosphate (15). The biological role of AcP is still uncertain, although there are some experimental indications as to its involvement in the regulation of membrane cation transport through the hydrolysis of phosphorylated intermediates formed during the action of membrane pumps (16,17).…”

mentioning

confidence: 99%

The Contribution of Acidic Residues to the Conformational Stability of Common-Type Acylphosphatase

Paoli

Taddei

Fiaschi

et al. 1999

Archives of Biochemistry and Biophysics

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Common-type acylphosphatase is a small cytosolic enzyme whose catalytic properties and three-dimensional structure are known in detail. All the acidic residues of the enzyme have been replaced by noncharged residues in order to assess their contributions to the conformational stability of acylphosphatase. The enzymatic activity parameters and the conformational free energy of each mutant were determined by enzymatic activity assays and chemically induced unfolding, respectively. Some mutants exhibit very similar conformational stability, ⌬G(H 2 O), and specific activity values as compared to the wild-type enzyme. By contrast, six mutants show a significant reduction of conformational stability and two mutants are more stable than the wild-type protein. Although none of the mutated acidic residues is directly involved in the catalytic mechanism of the enzyme, our results indicate that mutations of residues located on the surface of the protein are responsible for a structural distortion which propagate up to the active site. We found a good correlation between the free energy of unfolding and the enzymatic activity of acylphosphatase. This suggests that enzymatic activity measurements can provide valuable indications on the conformational stability of acylphosphatase mutants, provided the mutated residue lies far apart from the active site. Moreover, our results indicate that the distortion of hydrogen bonds rather than the loss of electrostatic interactions, contributes to the decrease of the conformational stability of the protein.

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Insights into acylphosphatase structure and catalytic mechanism

Cited by 84 publications

References 74 publications

The intrachain disulfide bridge is responsible of the unusual stability properties of novel acylphosphatase from Escherichia coli

The intrachain disulfide bridge is responsible of the unusual stability properties of novel acylphosphatase from Escherichia coli

Solution structure and conformational heterogeneity of acylphosphatase from Bacillus subtilis

The Contribution of Acidic Residues to the Conformational Stability of Common-Type Acylphosphatase

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