Effective expression and purification of recombinant onconase, an antitumor protein

FEBS Letters

Bracale, Aurora¹  D'Alessio, Giuseppe¹  Notomista, Eugenio¹  Di Donato, Alberto¹  Fusiello, Rossano¹  Cafaro, Valeria¹ 

[1] Dipartimentó di Chimica Organica e Biologica, Università di Napoli Federico II, Via Mezzocannone 16, 80134 Naples, Italy

关键词: Ribonuclease;    Onconase;    Antitumor;    Recombinant expression;    ONC;    onconase;    (M23L)-ONC;    mutant of onconase with a leucine residue at position 23 replacing methionine;    BS-RNase;    bovine seminal ribonuclease;    RNase A;    bovine pancreatic ribonuclease;    poly(U);    polyuridilic acid;    DTT;    dithiothreitol;    IPTG;    isopropyl-β-D-thiogalacto-pyranoside;    SDS-PAGE;    polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate;    CNBr;    cyanogen bromide;    ES-MS;    electrospray mass spectrometry;    TB;    terrific broth;    AAP;    Aeromonas proteolytica aminopeptidase;   

DOI  :  10.1016/S0014-5793(99)01623-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Several members of the RNase A superfamily are endowed with antitumor activity, showing selective cytotoxicity toward several tumor cell lines. One of these is onconase, the smallest member of the RNase A superfamily, which is at present undergoing phase III clinical trials. We report here the expression of recombinant onconase in Escherichia coli inclusion bodies, the correct processing of the protein, followed by its purification in high yields. The recombinant protein has biological and catalytic properties identical to those of the natural enzyme.

【 授权许可】

Unknown