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CAIII过氧化氢酶和过氧化物酶活性观察

发表时间:2016-09-15  浏览量:1244  下载量:522
全部作者: 尚西亮,陈疾忤,陈世益
作者单位: 复旦大学附属华山医院运动医学科
摘 要: 目的:观察骨骼肌碳酸酐酶III(carbonic anhydrase III,CAIII)是否具有过氧化氢酶或过氧化物酶活性。方法:提取骨骼肌可溶性蛋白,SDS-10%PAGE分离后分别进行考马斯亮蓝染色和Western blotting免疫染色,对照观察确定CAIII在凝胶上所处的位置;磷酸酶活性染色鉴定提取的CAIII是否仍然具有酶活性;铁染色和DAB显色法观察骨骼肌CAIII是否具有过氧化氢酶或过氧化物酶活性。结果:骨骼肌CAIII与抗CAIII多克隆抗体有较强的特异性反应条带,根据其位置可以在考马斯亮蓝染色图谱显示的复杂蛋白成分中鉴别出CAIII;酶活性染色表明提取的CAIII 仍然具有酶活性;铁染色法显示,在蓝绿色的凝胶染色背景下,CAIII相应位置未见有明显的透亮条带;DAB显色结果发现,在CAIII相应位置无明显的棕色条带。结论:CAIII可能不具有过氧化氢酶活性,也并非过氧化物酶同工酶,其抵御氧化应激损伤的机理有待于进一步研究。
关 键 词: 生物医学工程学;碳酸酐酶III;过氧化氢酶;过氧化物酶;磷酸酶
Title: Observation of catalase and peroxidase activity in carbonic anhydrase III
Author: SHANG Xiliang, CHEN Jiwu, CHEN Shiyi
Organization: Department of Sports Medicine, Huashan Hospital, Fudan University
Abstract: Objective: To observe whether carbonic anhydrase III (CAIII) in skeletal muscle has catalase or peroxidase activity. Methods: After extracting from skeletal muscle, the proteins were separated by means of SDS-10%PAGE. Kaumas blue staining and Western blotting staining were used to determine the location of CAIII on the gel. Phosphatase staining was used to identify if CAIII still had enzyme activity. Iron staining and DAB staining were used to observe whether CAIII had catalase or peroxidase activity. Results: CAIII in skeletal muscle could react immunologically with anti-CAIII polyclonal antibody. According to its location, CAIII could be identified from complex protein components on the Kaumas blue staining map. Phosphatase staining showed that CAIII still had enzyme activity. However, we did not find the corresponding CAIII strip by iron staining and DAB staining. Conclusion: CAIII may not have catalase activity, and it is also not a peroxidase isozyme. The mechanism of its resistence to oxidative damage needs to be further studied.
Key words: biomedical engineering; carbonic anhydrase III; catalase; peroxidase; phosphatase
发表期数: 2016年9月第17期
引用格式: 尚西亮,陈疾忤,陈世益. CAIII过氧化氢酶和过氧化物酶活性观察[J]. 中国科技论文在线精品论文,2016,9(17):1748-1752.
 
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