Michael Waterman
Faculty Member
Last active: 2/12/2015

Protein synthesis inhibitors and ethanol selectively enhance heterologous expression of P450s and related proteins in Escherichia coli.

Kusano K, Waterman MR, Sakaguchi M, Omura T, Kagawa N
Arch Biochem Biophys. 1999 367 (1): 129-36

PMID: 10375408 · DOI:10.1006/abbi.1999.1248

The antibiotics chloramphenicol (Cm), tetracycline, and erythromycin, which inhibit bacterial protein synthesis and are known to induce the cold shock response, unexpectedly enhance the heterologous expression of P450s and related proteins in Escherichia coli. In contrast, antibiotics that mimic heat shock in E. coli such as puromycin, streptomycin, and kanamycin decrease the expression of the same proteins. A sublethal dose of Cm (1 microgram/ml) effectively enhances the expression of both membrane-bound proteins (microsomal and mitochondrial P450s) and a soluble mitochondrial protein (adrenodoxin) over the range of two- to eightfold. The expression level of N-terminal truncated P450c17 (1600 nmol/liter culture without Cm), for instance, reached 3500 nmol/liter culture by the addition of Cm, approximately 8.4% of the total cellular protein. Cm also enabled expression at useful levels of active P450s previously difficult to express in E. coli. In contrast, the expression of P450scc, a mitochondrial protein, is decreased by Cm but enhanced by ethanol, a powerful elicitor of heat shock response in E. coli. These results show that both the cold shock response induced by some antibiotics and the heat shock response induced by ethanol may lead to enhanced expression of certain heterologous proteins in E. coli. This study also indicates that protein synthesis inhibitors associated with the cold shock response may act as protein synthesis enhancers under certain conditions.

Copyright 1999 Academic Press.

MeSH Terms (21)

Adrenodoxin Aminoglycosides Animals Anti-Bacterial Agents Cattle Chloramphenicol Cold Temperature Cytochrome P-450 Enzyme System Escherichia coli Ethanol Gene Expression Heat-Shock Response Isopropyl Thiogalactoside Membrane Proteins Microsomes Mitochondria Protein Synthesis Inhibitors Rats Recombinant Proteins Sequence Deletion Tetracycline

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