RT Journal Article
SR Electronic
T1 Effects in cancer cells of the recombinant L-Methionine gamma-lyase from Brevibacterium aurantiacum. Encapsulation in human erythrocytes for sustained L-methionine elimination
JF Journal of Pharmacology and Experimental Therapeutics
JO J Pharmacol Exp Ther
FD American Society for Pharmacology and Experimental Therapeutics
SP jpet.119.256537
DO 10.1124/jpet.119.256537
A1 David Machover
A1 Luigia Rossi
A1 Jocelyne Hamelin
A1 Christophe Desterke
A1 Emma Goldschmidt
A1 Bernadette Chadefaux-Vekemans
A1 Pascal Bonnarme
A1 Pierre Briozzo
A1 David Kopecny
A1 Francesca Pierige
A1 Mauro Magnani
A1 Rosella Mollicone
A1 Farhad Haghighi-Rad
A1 Yann Gaston-Mathe
A1 Julien Dairou
A1 Claude Boucheix
A1 Raphael Saffroy
YR 2019
UL http://jpet.aspetjournals.org/content/early/2019/04/02/jpet.119.256537.abstract
AB Methionine deprivation induces growth arrest and death of cancer cells. To eliminate L-methionine we produced, purified and characterized the recombinant pyridoxal 5'-phosphate (PLP)-dependent L-methionine gamma-lyase MGL-BL929 from the cheese-ripening Brevibacterium aurantiacum. Transformation of an Escherichia coli strain with the gene BL929 from B. aurantiacum optimized for E. coli expression led to production of the MGL-BL929. Elimination of L-methionine and cytotoxicity in vitro were assessed, and methylation-sensitive epigenetics was explored for changes resulting from exposure of cancer cells to the enzyme. A bioreactor was built by encapsulation of the protein in human erythrocytes to achieve sustained elimination of L-methionine in extracellular fluids. Catalysis was limited to α,γ-elimination of L-methionine and L-homocysteine. The enzyme had no activity on other sulfur-containing amino acids. Enzyme activity decreased in presence of serum albumin or plasma resulting from reduction of PLP availability. Elimination of L-methionine induced cytotoxicity on a vast panel of human cancer cell lines and spared normal cells. Exposure of colorectal carcinoma cells to the MGL-BL929 reduced methyl-CpG levels of hyper methylated gene promoters including that of CDKN2A whose mRNA expression was increased, together with decrease in global histone H3-dimethyl lysine9. The MGL erythrocyte bioreactor durably preserves enzyme activity in vitro and strongly eliminates L-methionine from medium.