Abstract: A DNA sequence coding for human muscle acylphosphatase has been constructed using 16 chemically synthesized oligonucleotides. The 300-bases long DNA sequence has been cloned in the pT7.7 Escherichia coli expression vector and in the pYEpsec1 Saccharomyces cerevisiae expression vector. In both cases a high level of expression of acylphosphatase has been observed. The recombinant proteins have been purified to homogeneity and assayed in comparison with the natural protein, using benzoylphosphate as a substrate and phosphate as a competitive inhibitor. The recombinant enzymes expressed in the two microorganisms maintain the kinetic properties of the natural protein. In addition, NMR analysis shows that the gross fold of the two recombinant enzymes is correct.

Chemical synthesis and expression of a gene coding for human muscle acylphosphatase / A. Modesti; G. Raugei; N. Taddei; R. Marzocchini; M. Vecchi; G. Camici; G. Manao; G. Ramponi. - In: BIOCHIMICA ET BIOPHYSICA ACTA. - ISSN 0006-3002. - STAMPA. - 1216:(1993), pp. 369-374. [WOS:A1993MP36600003]

Chemical synthesis and expression of a gene coding for human muscle acylphosphatase

MODESTI, ALESSANDRA;RAUGEI, GIOVANNI;TADDEI, NICCOLO';MARZOCCHINI, RICCARDO;CAMICI, GUIDO;MANAO, GIAMPAOLO;RAMPONI, GIAMPIETRO
1993

Abstract

Abstract: A DNA sequence coding for human muscle acylphosphatase has been constructed using 16 chemically synthesized oligonucleotides. The 300-bases long DNA sequence has been cloned in the pT7.7 Escherichia coli expression vector and in the pYEpsec1 Saccharomyces cerevisiae expression vector. In both cases a high level of expression of acylphosphatase has been observed. The recombinant proteins have been purified to homogeneity and assayed in comparison with the natural protein, using benzoylphosphate as a substrate and phosphate as a competitive inhibitor. The recombinant enzymes expressed in the two microorganisms maintain the kinetic properties of the natural protein. In addition, NMR analysis shows that the gross fold of the two recombinant enzymes is correct.
1993
1216
369
374
A. Modesti; G. Raugei; N. Taddei; R. Marzocchini; M. Vecchi; G. Camici; G. Manao; G. Ramponi
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/323410
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