Biochemical characteristics ofS-adenosylmethionine decarboxylase from carnation (Dianthus caryophyllus L.) petals |
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Authors: | Myeong Min Lee Kyoung Hee Nam Eun Kyoung Lee Sun Hi Lee Ky Young Park |
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Affiliation: | (1) Department of Biology, Yonsei University, Sinchon 134, 120-749 Seoul, Korea;(2) Department of Biology, Sunchon National University, 540-742 Sunchon, Chonnam, Korea |
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Abstract: | We have partially purified S-adenosylmethionine decarboxylase (EC 4.1.1.50, SAMDC) from carnation (Dianthus caryophyllus L.) petals and generated polyclonal antibodies against CSDC 16 protein (Leeet al., 1996) overexpressed inE. coli. The protein has been purified approximately 126.8 fold through the steps involving ammonium sulfate fractionation, DEAE-Sepharose column chromatography and Sephacryl S-300 gel filtration. Its molecular mass was 42 kDa in native form and we could also detect a band of 32 kDa molecular mass on SDS-PAGE in western blot analysis using the polyclonal antibodies. The Km value of this enzyme forS-adenosylmethionine was 26.3 μM. The optimum temperature and pH forS-adenosylmethionine decarboxylase activity were 35°C and pH 8.0, respectively. Putrescine and Mg2+ had no effects on the activation of the enzyme activity. Mg2+ did not have any significant effects on the enzyme activity. SAMDC activity was inhibited by putrescine, spermidine and spermine. Methylglyoxal bis-(guanylhydrazone) (MGBG), carbonyl reagents such as hydroxylamine and phenylhydrazine, and sulfhydryl reagent such as 5,5′dithio-bis (2-nitrobenzoic acid) (DTNB) were effective inhibitors of the enzyme. However, isonicotinic acid hydrazide known as an inhibitor of 5′-pyridoxal phosphate (PLP) dependent enzyme activity had no significant effect on the enzyme activity. These results and our previously reported results (Leeet al., 1997b) suggest thatS-adenosylmethionine decarboxylase is a heterodimer, αβ, and some carbonyl group and sulfhydryl group are involved in the catalytic activity. |
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Keywords: | SAMDC biochemical characteristic molecular mass antibody processing |
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