Schweiger A, Kostka G
Biochim Biophys Acta. 1985 Nov 13;826(2-3):87-94. doi: 10.1016/0167-4781(85)90112-5.
Isolated liver nuclei or whole lymph node lymphocytes stimulated with concanavalin A in culture were irradiated with ultraviolet light. The crosslinked structures of poly(A)+ heterogeneous nuclear RNA and protein were purified on oligo(dT)-cellulose after labelling irradiated nuclei in the presence of adenosine 5'-[gamma-32P]triphosphate and analysed by SDS-polyacrylamide gel electrophoresis. The liver and lymphocyte nuclear proteins included about 17-19 species of 35-150 kDa and were shown to produce quite similar electrophoretic band patterns. Two proteins of 110-120 and 40-42 kDa were phosphorylated. Using partial proteolytic digestion the large-size crosslinked phosphoprotein has been identified as the 110 kDa component described previously (Schweiger, A. and Kostka, G. (1984) Biochim. Biophys. Acta 782, 262-268). The 40-42 kDa band was presumably related to the group C species of main proteins associated with heterogeneous nuclear RNA. In crosslinked nuclear structures from rats treated with low doses of alpha-amanitin for 1 h the relative amount of the 110-120 kDa phosphoprotein was reduced while the labelling with [32P]ATP was almost abolished.
将培养中用伴刀豆球蛋白A刺激的分离肝细胞核或全淋巴结淋巴细胞用紫外线照射。在用5'-[γ-32P]三磷酸腺苷存在下标记受照射的核后，在寡聚（dT）-纤维素上纯化聚（A）+异质核RNA和蛋白质的交联结构，并通过SDS-聚丙烯酰胺凝胶电泳进行分析。肝和淋巴细胞核蛋白包括约17-19种35-150 kDa的蛋白，并且显示出产生相当相似的电泳条带模式。两种110-120 kDa和40-42 kDa的蛋白被磷酸化。通过部分蛋白酶解消化，已将大尺寸交联磷蛋白鉴定为先前描述的110 kDa组分（施魏格，A.和科斯特卡，G.（1984年）生物化学与生物物理学报782，262-268）。40-42 kDa条带可能与与异质核RNA相关的主要蛋白的C组物种有关。在用低剂量α-鹅膏蕈碱处理1小时的大鼠的交联核结构中，110-120 kDa磷蛋白的相对量减少，而用[32P]ATP的标记几乎被消除。
