title: Catalase activity is modulated by calcium and calmodulin in detached mature leaves of sweet potato.
abstract: Catalase (CAT) functions as one of the key enzymes in the scavenging of reactive oxygen species and affects the H2O2 homeostasis in plants. In sweet potato, a major catalase isoform was detected, and total catalase activity showed the highest level in mature leaves (L3) compared to immature (L1) and completely yellow, senescent leaves (L5). The major catalase isoform as well as total enzymatic activity were strongly suppressed by ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA). This inhibition could be specifically and significantly mitigated in mature L3 leaves by exogenous CaCl2, but not MgCl2 or CoCl2. EGTA also inhibited the activity of the catalase isoform in vitro. Furthermore, chlorpromazine (CPZ), a calmodulin (CAM) inhibitor, drastically suppressed the major catalase isoform as well as total enzymatic activity, and this suppression was alleviated by exogenous sweet potato calmodulin (SPCAM) fusion protein in L3 leaves. CPZ also inhibited the activity of the catalase isoform in vitro. Protein blot hybridization showed that both anti-catalase SPCAT1 and anti-calmodulin SPCAM antibodies detect a band at the same position, which corresponds to the activity of the major catalase isoform from unboiled, but not boiled crude protein extract of L3 leaves. An inverse correlation between the major catalase isoform/total enzymatic activity and the H2O2 level was also observed. These data suggest that sweet potato CAT activity is modulated by CaCl2 and SPCAM, and plays an important role in H2O2 homeostasis in mature leaves. Association of SPCAM with the major CAT isoform is required and regulates the in-gel CAT activity band.
authors: Department of Biological Sciences, National Sun Yat-sen University, 80424 Kaohsiung, Taiwan.
authors: Mufidah
authors: M
authors: Afiyanti
authors: Department of Biological Sciences, National Sun Yat-sen University, 80424 Kaohsiung, Taiwan. Electronic address: hjchen@faculty.nsysu.edu.tw.
authors: Hsien-Jung
authors: HJ
authors: Chen
mesh_terms: D002122:Calcium Chloride / Q000737:chemistry* / Q000378:metabolism; D002147:Calmodulin / Q000737:chemistry* / Q000378:metabolism; D002374:Catalase / Q000378:metabolism*; D004533:Egtazic Acid; D006861:Hydrogen Peroxide / Q000378:metabolism*; D027723:Ipomoea batatas / Q000201:enzymology*; D007527:Isoenzymes / Q000378:metabolism; D018515:Plant Leaves / Q000378:metabolism
chemical_list: D002147:Calmodulin; D007527:Isoenzymes; D004533:Egtazic Acid; D006861:Hydrogen Peroxide; D002374:Catalase; D002122:Calcium Chloride
keywords: CAM; CAT; CCI; CPZ; Calcium; Calmodulin; Catalase; Chlorpromazine; EGTA; SDS-PAGE; SPCAM; Sweet potato; calmodulin; catalase; chlorophyll content index; chlorpromazine; ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid; sodium dodecyl sulfate-polyacrylamide gel electrophoresis; sweet potato calmodulin