Efficient production of active TNF‐α by albumin signal peptide

TNF‐α is initially synthesized as a membrane‐anchored precursor protein and processed proteolytically by a matrix metalloproteinase (MMP)‐like enzyme. In order to establish an efficient expression system of TNF‐α in mammalian cells without involvement of the extracellular enzyme, an expression plasm...

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Bibliographic Details
Published in:Biochemistry and molecular biology international 1997-07, Vol.42 (4), p.825-832
Main Authors: Maeda, Yuu, Soda, Mariko, Ito, Keizo, Sato, Kenzo
Format: Article
Language:English
Subjects:
recombinant protein
secretion
signal peptide
TNF‐α
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Summary:TNF‐α is initially synthesized as a membrane‐anchored precursor protein and processed proteolytically by a matrix metalloproteinase (MMP)‐like enzyme. In order to establish an efficient expression system of TNF‐α in mammalian cells without involvement of the extracellular enzyme, an expression plasmid (pCN‐alb‐TNF) was constructed with a signal sequence of the rat albumin gene as a module for secretion. The highest level of production of TNF‐α was observed in the clone CT‐3 by SDS‐PAGE and Western blot analysis. Biological activity of the secretion was revealed by repression of catalase gene expression in hepatoma cells and cytotoxity to L929 cells. Attachment of the signal peptide to mature form resulted in the enhancement of production of the cytokine.
ISSN:1521-6543
1039-9712
1521-6551
DOI:10.1080/15216549700203261