Kyriakopoulos A, Röthlein D, Pfeifer H, Bertelsmann H, Kappler S, Behne D
Hahn-Meitner-lnstitut Berlin, Department "Trace Elements in Health and Nutrition", Germany.
J Trace Elem Med Biol. 2000 Oct;14(3):179-83. doi: 10.1016/S0946-672X(00)80008-5.
The important role of selenium in the mammalian organism has been manifested by the detection of several selenoenzymes, and there are still numerous selenium-containing proteins to be identified. After in vivo labeling of rats with [75Se]-selenite, gel electrophoretic separation of the proteins in tissue homogenates and autoradiography of the labeled bands, information on the selenium-containing proteins present in the different tissues was obtained. In the separation by SDS-PAGE and two-dimensional IEF/SDS-PAGE a large number of selenium-containing proteins or protein subunits with apparent molecular masses in the range from 116 to 8 kDa could be distinguished. This range was extended by applying a modified Tricine-SDS-PAGE, which allows the determination of smaller proteins. Using this method in the separation of the homogenates of the adrenal, brain, diaphragm, epididymis, heart, kidney, liver, lung, pituitary, prostate, skeletal muscle, spleen, thymus and thyroid, four additional selenium-containing proteins with molecular masses of approximately 7 kDa, 5kDa, 4 kDa and 3kDa were detected. The 5 kDa protein and the 7 kDa protein were identified as selenocysteine-containing selenoproteins.
通过几种含硒酶的发现，硒在哺乳动物机体中的重要作用得以体现，并且仍有众多含硒蛋白质有待鉴定。在用[75Se] -亚硒酸盐对大鼠进行体内标记后，对组织匀浆中的蛋白质进行凝胶电泳分离以及对标记条带进行放射自显影，从而获得了不同组织中含硒蛋白质的相关信息。在通过SDS - PAGE和二维IEF/SDS - PAGE进行分离时，可以区分出大量表观分子量在116至8 kDa范围内的含硒蛋白质或蛋白质亚基。通过应用改良的Tricine - SDS - PAGE扩展了这个范围，该方法能够测定更小的蛋白质。使用这种方法分离肾上腺、脑、横膈膜、附睾、心脏、肾脏、肝脏、肺、垂体、前列腺、骨骼肌、脾脏、胸腺和甲状腺的匀浆时，检测到另外四种分子量约为7 kDa、5 kDa、4 kDa和3 kDa的含硒蛋白质。5 kDa蛋白质和7 kDa蛋白质被鉴定为含硒代半胱氨酸的硒蛋白。
