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Comparison of Calpains from Rabbit, Monkey, Human and Rat

机译:兔,猴,人和大鼠中钙蛋白酶的比较

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Two isozymes of calpain, μ-calpain and m-calpain, were purified from rabbit, monkey, human and rat tissues to homogeneity and the apparent molecular masses of the large and small subunits of each calpain species were compared directly. While the molecular masses of the small subunits were the same (28 kDa), those of the large subunits were different depending on the calpain type and animal species: Rabbit μ (79 kDa), rabbitm (75 kDa), monkey μ (79 kDa), monkey m (74 kDa), human μ (78 kDa), human m (73 kDa), rat μ(75 kDa), and rat m (74 kDa). Ca~(2+)-sensitivity of monkey μ-calpain was lower than that of rabbit μ-calpain, but m-calpains from rabbit and monkey shared a similar Ca~(2+)-dependency. Immunoreactivities of rabbit, monkey and rat m-calpains towards anti-rabbit m-calpain monoclonal antibodies were different depending on the antibody species, showing the existence of common and different antigenic sites in these three m-calpains. While monkey μ-calpain still showed weak cross-reactivity with anti-rabbit μ-calpain monoclonal antibodies, rat μ-calpain failed to react with any antibody examined, except for the monoclonal 1D10A7 which reacted with μ- and m-calpains from any animal species. Peptides generated by V8 protease digestion were similar between μ-calpains or m-calpains from rabbit and monkey but, again, the reactivity of monkey calpain peptides to anti-rabbit calpain antibodies was weak, especially to those of μ-calpain.
机译:从兔,猴,人和大鼠组织中纯化出钙蛋白酶的两种同工酶μ-钙蛋白酶和间钙蛋白酶,以达到均质,并直接比较每种钙蛋白酶物种的大,小亚基的表观分子量。虽然小亚基的分子质量相同(28 kDa),但大亚基的分子质量因钙蛋白酶类型和动物种类而异:兔子μ(79 kDa),兔子(75 kDa),猴子μ(79 kDa) ),猴子m(74 kDa),人μ(78 kDa),人m(73 kDa),大鼠μ(75 kDa)和大鼠m(74 kDa)。猴子μ-钙蛋白酶对Ca〜(2+)的敏感性低于兔子μ-钙蛋白酶,但来自兔子和猴子的m-钙蛋白酶具有相似的Ca〜(2+)依赖性。兔,猴子和大鼠的m-钙蛋白酶对抗兔m-钙蛋白酶单克隆抗体的免疫反应性因抗体种类而异,表明这三种m-钙蛋白酶存在相同和不同的抗原位点。尽管猴子μ-钙蛋白酶与抗兔μ-钙蛋白酶单克隆抗体的交叉反应性仍然很弱,但大鼠μ-钙蛋白酶不能与所检测的任何抗体反应,除了单克隆1D10A7可以与任何动物的μ-钙蛋白酶和m-钙蛋白酶反应之外种类。 V8蛋白酶消化产生的肽在兔和猴子的μ-钙蛋白酶或m-钙蛋白酶之间相似,但再次,猴子钙蛋白酶肽对抗兔钙蛋白酶抗体的反应性较弱,特别是对μ-钙蛋白酶。

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