Expression, purification, and characterization of acylphosphatase muscular isoenzyme as fusion protein with glutathione S-transferase.
Protein Expr Purif
; 6(6): 799-805, 1995 Dec.
Article
em En
| MEDLINE
| ID: mdl-8746632
ABSTRACT
A genetic construct consisting of the synthetic gene coding for human muscle acylphosphatase linked to the gene for glutathione S-transferase has been prepared. This gene was transformed into and expressed by the Escherichia coli strains DB1035 and TB1, respectively. The fusion protein was purified by affinity chromatography and subsequently cleaved to the fully active acylphosphatase, which was further purified by gel filtration chromatography. Such a purification procedure is very rapid and suitable for obtaining considerable amounts of enzyme at a very high yield. The purified human muscle acylphosphatase was fully active and showed structural features, as well as kinetic and stability parameters, identical to those of the native enzyme.
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Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Hidrolases Anidrido Ácido
/
Isoenzimas
/
Músculos
Limite:
Humans
Idioma:
En
Revista:
Protein Expr Purif
Ano de publicação:
1995
Tipo de documento:
Article