Leucine Aminopeptidase from Red Sea Bream (Pagrus major) Skeletal Muscle: Purification, Characterization, Cellular Location, and Tissue Distribution | |
Su, Wen-Jin ; Su WJ(苏文金) ; Wu, Guo-Ping ; Cao, Min-Jie(Biol Engn, Xiamen) ; Liu, Bing-Xin(Biol Engn, Xiamen) ; Chen, Yan(Jimei Univ) | |
2008 | |
关键词 | Red sea bream leucine aminopeptidase purification characterization Western blot |
英文摘要 | A leucine aminopeptidase was purified for the first time from marine fish red sea bream (Pagrus major skeletal muscle to homogeneity with 4850-fold and a yield of 7.4%. The purification procedure consisted of ammonium sulfate fractionation and chromatographies including DEAE-Sephacel, Sephacryl S-200, hydroxyapatite, and phenyl-Sepharose. The enzyme was approximately 96 kDa as estimated by SIDS-PAGE and gel filtration and preferentially hydrolyzed substrate Leu-MCA. The enzymatic activity was optimal at 45 degrees C and pH 7.5. The K-m and k(cat) values of the enzyme for Leu-MCA were 1,55 mu M and 26.4 S-1 at 37 degrees C, respectively. Activation energy (E-a) of the enzyme was 59.6 kJ M-1. The enzyme was specifically inhibited by metal-chelating agents, and Zn2+ and (or) Mn2+ seemed to be its metal cofactor(s). In addition, bestatin strongly inhibited its activity, and K-i was 1.44 mu M. Using a highly specific polyclonal antibody, the location of enzyme was demonstrated intracellularly and distributed in different tissues.; National Natural Scientific Foundation of China [30571450]; Natural Scientific Foundation of Fujian Province [2006J0368]; Foundation for Innovative Research Team of Jimei University [2006A002] |
语种 | 英语 |
出版者 | AMER CHEMICAL SOC |
内容类型 | 期刊论文 |
源URL | [http://dx.doi.org/doi:10.1021/jf801477r] ![]() |
专题 | 生命科学-已发表论文 |
推荐引用方式 GB/T 7714 | Su, Wen-Jin,Su WJ,Wu, Guo-Ping,et al. Leucine Aminopeptidase from Red Sea Bream (Pagrus major) Skeletal Muscle: Purification, Characterization, Cellular Location, and Tissue Distribution[J],2008. |
APA | Su, Wen-Jin,苏文金,Wu, Guo-Ping,Cao, Min-Jie,Liu, Bing-Xin,&Chen, Yan.(2008).Leucine Aminopeptidase from Red Sea Bream (Pagrus major) Skeletal Muscle: Purification, Characterization, Cellular Location, and Tissue Distribution.. |
MLA | Su, Wen-Jin,et al."Leucine Aminopeptidase from Red Sea Bream (Pagrus major) Skeletal Muscle: Purification, Characterization, Cellular Location, and Tissue Distribution".(2008). |
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