Watanabe K, Akiyama K, Hikichi K, Ohtsuka R, Okuyama A, Senshu T
Tokyo Metropolitan Institute of Gerontology, Japan.
Biochim Biophys Acta. 1988 Sep 8;966(3):375-83. doi: 10.1016/0304-4165(88)90088-8.
We have performed a combined biochemical and immunochemical study on the identity of peptidylarginine deiminases (EC 3.5.3.15) present in various mammalian tissues. First, we purified peptidylarginine deiminase from rat skeletal muscle. It gave a single band of molecular weight 83,000 in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Next we immunized rabbits with the purified enzyme. The resulting antibodies reacted specifically with the antigen in Western blot assay. Most of the enzyme activities present in rat skeletal muscle, brain, spinal cord, submaxillary gland and spleen could be characterized as the same muscle-type enzyme by immunoprecipitation and Western blot assay. The antibodies did not react with enzyme samples obtained from rat hair follicles and bovine epidermis. The lack of immunoreactivity of the epidermal enzyme could not be accounted for by the species difference, since the antibodies reacted with a 83 kDa polypeptide of bovine brain, which was thought to represent a bovine counterpart of the muscle-type enzyme. The epidermal enzyme could be distinguished from the other enzyme samples by its high activity towards benzoylarginine. These data suggest the existence of at least three types of peptidylarginine deiminase in mammalian tissues, i.e., a muscle type, a hair follicle type, and an epidermal type.
我们对多种哺乳动物组织中肽基精氨酸脱亚氨酶（EC 3.5.3.15）的特性进行了生物化学与免疫化学联合研究。首先，我们从大鼠骨骼肌中纯化了肽基精氨酸脱亚氨酶。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中，它呈现出一条分子量为83,000的单一条带。接下来，我们用纯化后的酶免疫兔子。所得抗体在蛋白质印迹分析中与抗原发生特异性反应。通过免疫沉淀和蛋白质印迹分析，大鼠骨骼肌、脑、脊髓、下颌下腺和脾脏中存在的大部分酶活性可被鉴定为同一种肌肉型酶。这些抗体与从大鼠毛囊和牛表皮获得的酶样品不发生反应。表皮酶缺乏免疫反应性不能用物种差异来解释，因为这些抗体与牛脑的一种83 kDa多肽发生反应，该多肽被认为是肌肉型酶的牛对应物。表皮酶可通过其对苯甲酰精氨酸的高活性与其他酶样品区分开来。这些数据表明哺乳动物组织中至少存在三种类型的肽基精氨酸脱亚氨酶，即肌肉型、毛囊型和表皮型。
