New muteins of RNase A with enhanced antitumor action

FEBS Letters

Bracale, Aurora¹  D'Alessio, Giuseppe¹  Sorrentino, Salvatore¹  Di Maro, Antimo¹  Di Donato, Alberto¹  Cafaro, Valeria¹ 

[1] Department of Organic and Biological Chemistry, University of Naples Federico II, Via Mezzocannone 16, 80134 Naples, Italy

关键词: Ribonuclease;    Antitumor;    Protein engineering;    RNase A;    bovine pancreatic ribonuclease A;    BS-RNase;    bovine seminal RNase;    PLCC-RNase AA;    (A19P;    Q28L;    K31C;    S32C) dimeric RNase A;    PLCC(G38)-RNase AA;    (A19P;    Q28L;    K31C;    S32C;    D38G) dimeric RNase A;    PLCC(G111)-RNase AA;    (A19P;    Q28L;    K31C;    S32C;    E11G) dimeric RNase A;    PLCCGG-RNase AA;    (A19P;    Q28L;    K31C;    S32C;    G38;    G111) dimeric RNase A;    EDTA;    ethylenediamine tetraacetic acid;    PAGE;    polyacrylamide gel electrophoresis;    SDS;    sodium dodecylsulfate;    Tris;    tris(hydroxymethyl)aminomethane;    DTT;    dithiothreitol;    IPTG;    isopropyl-β-d-thiogalactopyranoside;    MOPS;    morpholinepropane sulfonic acid;    Poly(A)·Poly(U);    polyadenylic-polyuridylic acid (double-stranded);    dsRNA;    double stranded RNA;   

DOI  :  10.1016/S0014-5793(98)01221-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Monomeric bovine pancreatic RNase A has been transformed into a dimeric ribonuclease with antitumor activity (Di Donato, A., Cafaro, V. and D'Alessio, G. (1994) J. Biol. Chem. 269, 17394–17396). This was accomplished by replacing the residues located in the RNase chain at positions 19, 28, 31, and 32, with proline, leucine, and two cysteine residues, respectively, i.e. those present at identical positions in the subunit of bovine seminal RNase, a dimeric RNase of the pancreatic-type superfamily, endowed with a powerful antitumor action. However, as an antitumor agent this mutant dimeric RNase A is not as powerful as seminal RNase. We report here site-directed mutagenesis experiments which have led to the identification of two other amino acid residues, glycine 38 and 111, whose substitution in the polypeptide chain of the first generation dimeric mutant of RNase A, is capable of conferring to the mutein the full cytotoxic activity characteristic of native seminal RNase.

【 授权许可】

Unknown