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The intrachain disulfide bridge is responsible of the unusual stability properties of novel acylphosphatase from Escherichia coli
Author(s) -
Ramazzotti Matteo,
Parrini Claudia,
Stefani Massimo,
Manao Giampaolo,
Degl'Innocenti Donatella
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2006.11.033
Subject(s) - escherichia coli , mutagenesis , biochemistry , open reading frame , recombinant dna , chemistry , enzyme , site directed mutagenesis , gene , disulfide bond , thermophile , biology , peptide sequence , mutation , mutant
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.