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.

The intrachain disulfide bridge is responsible of the unusual stability properties of novel acylphosphatase from Escherichia coli / M. Ramazzotti; C. Parrini; M. Stefani; G. Manao; D. Degl'Innocenti. - In: FEBS LETTERS. - ISSN 0014-5793. - STAMPA. - 580:(2006), pp. 6763-6768. [10.1016/j.febslet.2006.11.033]

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

RAMAZZOTTI, MATTEO;STEFANI, MASSIMO;MANAO, GIAMPAOLO;DEGL'INNOCENTI, DONATELLA
2006

Abstract

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.
2006
580
6763
6768
M. Ramazzotti; C. Parrini; M. Stefani; G. Manao; D. Degl'Innocenti
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/322197
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