title: Expression of catalytically active rat S-adenosylmethionine decarboxylase in Escherichia coli.
abstract: The cDNA coding for rat S-adenosylmethionine decarboxylase (AdoMetDC, EC 4.1.1.50) has been cloned into a plasmid expression vector, pKK-223-3, and expressed in E. coli. The authenticity of the expressed protein has been demonstrated by reactivity with antibodies specific for rat AdoMetDC, by size analysis on SDS gels visualized with immunotransblots, and, finally, by catalytic activity. The expression of the enzyme results in a decrease in the activity of the bacterial enzyme suggesting the replacement of the bacterial enzyme by the rat AdoMetDC. Similarly, the addition of exogenous spermidine to the growth medium reduces bacterial enzyme activity affecting only marginally the expression of the recombinant protein.
authors.affiliation: Biocenter, University of Oulu, Finland.
authors.forename: P
authors.initials: P
authors.lastname: Salmikangas
authors.forename: M R
authors.initials: MR
authors.lastname: Keränen
authors.forename: A
authors.initials: A
authors.lastname: Pajunen
mesh_terms: D012437:Adenosylmethionine Decarboxylase / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D002262:Carboxy-Lyases / Q000235:genetics*; D002384:Catalysis; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism; D010957:Plasmids; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D013095:Spermidine / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; D004247:DNA; D002262:Carboxy-Lyases; D012437:Adenosylmethionine Decarboxylase; D013095:Spermidine