Polyamine synthesis in plants. Purification and properties of amidinotransferase from soybean (Glycine max) axes.
Three-day-old soybean (Glycine max) seedlings were exposed to 0.4 M sorbitol solution for 4 h to induce amidinotransferase activity, with the corresponding enzyme being purified to homogeneity by chromatographic separation on DEAE-Sephacel, Sephacryl S-300 and L-arginine Sepharose 4B. The purified enzyme used L-arginine and L-glycine as the major donor/acceptor of the amidino group, respectively, with formation of guanidinoacetic acid and ornithine products being confirmed by ESI-MS. The enzyme is a tetrameric protein having a molecular mass of 240,000 Da, whose thiol group is needed for enzymatic activity. The K(M)s for arginine and glycine were 3.8 and 0.89 mM, respectively, with optimal temperature and pH being 37 degrees C and 9.5, respectively. The soybean amidinotransferase could be indirectly involved in nitrogen metabolism, as suggested by the observation that arginine:glycine amidinotransferase in soybean axes is indirectly involved in putrescine biosynthesis and displays feedback control at high levels of an endogenous regulator, putrescine.
Lee
Geun Taek
GT
Department of Biochemistry, Yonsei University, 134 Shinchon-Dong, Seodaemoon-Ku, Seoul, South Korea.
Kim
Woo Jeung
WJ
Cho
Young Dong
YD
D000579:Amidinotransferases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D001120:Arginine / Q000737:chemistry / Q000378:metabolism; D015317:Biogenic Polyamines / Q000096:biosynthesis* / Q000378:metabolism; D002848:Chromatography, DEAE-Cellulose; D004789:Enzyme Activation / Q000187:drug effects; D005998:Glycine / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D013012:Sorbitol / Q000494:pharmacology; D013025:Glycine max / Q000201:enzymology*; D021241:Spectrometry, Mass, Electrospray Ionization; D013379:Substrate Specificity
D015317:Biogenic Polyamines; D013012:Sorbitol; D001120:Arginine; D000579:Amidinotransferases; D005998:Glycine