Expression in mammalian cells, purification and characterization of recombinant human pancreatic ribonuclease

FEBS Letters

Ciardiello, Antonietta¹  de Nigris, Mena¹  D'Alessio, Giuseppe¹  Di Donato, Alberto¹  Russo, Nello¹ 

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

关键词: Ribonuclease;    Human pancreatic RNase;    expression;    HP-RNase;    human pancreatic ribonuclease;    RNase A;    bovine pancreatic ribonuclease A;    RISBASES;    ribonucleases with special biological actions;    CHO cells;    Chinese hamster ovary cells;    poly(A):poly(U);    double-stranded polymer made up of poly-AMP and of poly-UMP;    SDS-PAGE;    polyacrylamide gel electrophoresis in sodium dodecyl sulfate;   

DOI  :  10.1016/0014-5793(93)80660-M

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A synthetic cDNA coding for human pancreatic RNase, equipped with a secretion signal sequence, was cloned and stably expressed in Chinese hamster ovary cells. The recombinant RNase, secreted into the culture medium, was purified and characterized. It was found to be indistinguishable, by structural and catalytic parameters, from the enzyme isolated from human pancreas. Furthermore, the glycosylated forms were separated from the non-glycosylated form. Up until now, human RNases have been isolated only in small amounts from autoptic specimens. This has hindered the exploitation of a human RNase for the construction of immunotolerated immunotoxins. On the other hand, the availability of an effective system for the expression of a human RNase may render feasible the transfer, by protein engineering, of the interesting pharmacological actions of non-human RNase [1993 Trends Cell Biol. 3, 106-109] to an immunotolerated, human RNase.

【 授权许可】

Unknown