Novel immunoassays to detect methionine adenosyltransferase activity and quantify S-adenosylmethionine

Abstract

We present a novel real-time immunoassay to measure methionine adenosyltransferase (MAT) activity that integrates the MAT-catalyzed reaction of Met and adenosine triphosphate to produce S-adenosylmethionine (SAM) and a highly sensitive immunoassay to specifically quantify SAM simultaneously. The cellular localization of SAM and S-adenosylhomocysteine varies with cell proliferation status: in normal cells, they are found mostly in the cytoplasm, but localize to the nucleus in proliferating cells. MAT-I/III activity is stimulated by Met, but inhibited by S-nitrosoglutathione, and the methylation index (MI) increases after Met stimulation of L02 cells. Met and S-nitrosoglutathione inhibit MAT-II activity, and the MI decreases after Met stimulation of HepG2 cells. The method described provides a significant advancement in the field for the measurement of MAT activity under various conditions.