The pro-sequence domain of streptopain directs the folding of the mature enzyme.
Arch Biochem Biophys
; 436(2): 297-306, 2005 Apr 15.
Article
em En
| MEDLINE
| ID: mdl-15797242
ABSTRACT
The cysteine endopeptidase streptopain, an extracellular enzyme from pathogenic Streptococcus pyogenes, is synthesized as a precursor containing an NH2-terminal pro-sequence. The pro-sequence of streptopain was expressed in Escherichia coli and subjected to structural and functional investigation. Heat-induced denaturation of the pro-sequence studied using circular dichroism spectroscopy revealed that it forms a compact structure and represents an independently folded domain. The isolated pro-sequence exhibits high affinity towards mature streptopain and associates with its cognate enzyme by forming an equimolar complex. Refolding of denatured streptopain in the presence of pro-sequence in vitro facilitated recovery of active enzyme. Expression of the mature streptopain in E. coli either alone, or in trans with its pro-sequence as an independent polypeptide, led to the formation of insoluble protein aggregates or functionally active enzyme, respectively. These results demonstrate that the pro-sequence domain acts as an intramolecular chaperone that directs the correct folding of the mature streptopain.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Streptococcus pyogenes
/
Cisteína Endopeptidases
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
Arch Biochem Biophys
Ano de publicação:
2005
Tipo de documento:
Article
País de afiliação:
Estados Unidos