Expression of dipeptidyl peptidase III in normal lenses of rat of different ages

Haiou An, Hui Zhang, Xiu-Yuan Qin, Rui Tian, Ying Dou, Xuexun Fang
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Abstract

Background It has been determined that dipeptidyl peptidase Ⅲ (DPPⅢ) plays an important role in the metabolism and modification of proteins and DPPⅢ of human has highly homologous to rat.Researches have shown that DPPⅢ is associated with the formation of cataract.However,few relevant studies have been reported.ObjectiveThe present study is to find out the relationship between the expression of DPPⅢ in rat lenses and age-related cataract.Methods Lenses were obtained from general Wistar rats of ages 3,6,9,or 12 weeks old (10 lenses each ) and homogenized with different concentrations of standard bovine serum.The proteins were resolved using SDS-PAGE gel electrophoresis.Peak concentration and total concentration of DPPⅢ protein in lenses from rats of different ages were detected by Western blot.Enzyme activity of DPPⅢ was determined by monitoring the amount of dipeptides removed from a special substrate (Arg-Arg-4mb NA) by measuring the absorbance with UV-2500PC at 525 nm.The relationship between the enzyme activity of DPPⅢ in lenses and age of rats was evaluated using regression analysis.Results DPPⅢ was detected at a molecular weight of 82000 Da.The peak concentration and total concentration of DPPⅢ in normal rat lenses increased with the growth of age.The total protease activity of DPPⅢ in the lenses of rats was correlated with the ages of the rats (r=0.99,P<0.05).Conclusion DPPⅢ may be involved in the alteration of crystallin during the development of lenses,and it may play an important role in the formation and aggravation of age-related cataract. Key words: dipeptidyl peptidase Ⅲ; lens; age-related cataract; enzymes activity
二肽基肽酶III在不同年龄大鼠正常晶状体中的表达
二肽基肽酶Ⅲ(DPPⅢ)在蛋白质的代谢和修饰中起着重要的作用,人的DPPⅢ与大鼠具有高度的同源性。研究表明DPPⅢ与白内障的形成有关。然而,相关研究鲜有报道。目的探讨大鼠晶状体中DPPⅢ表达与老年性白内障的关系。方法取3、6、9、12周龄Wistar大鼠(每只10个)晶状体,用不同浓度的标准牛血清均质。采用SDS-PAGE凝胶电泳对蛋白进行分离。Western blot法检测不同年龄大鼠晶状体中DPPⅢ蛋白的峰值浓度和总浓度。用UV-2500PC在525 nm处测量吸光度,监测从特殊底物(Arg-Arg-4mb NA)上去除的二肽的量,从而测定DPPⅢ的酶活性。采用回归分析评价大鼠晶状体DPPⅢ酶活性与年龄的关系。结果检测到分子量为82000 Da的DPPⅢ。正常大鼠晶状体中DPPⅢ的峰值浓度和总浓度随着年龄的增长而增加。大鼠晶状体DPPⅢ总蛋白酶活性与大鼠年龄相关(r=0.99,P<0.05)。结论DPPⅢ可能参与晶状体发育过程中晶状体蛋白的改变,在老年性白内障的形成和加重中起重要作用。关键词:二肽基肽酶Ⅲ;镜头;年龄相关性白内障;酶的活动
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