Ohmura Y, Teraoka H, Tsukada K
FEBS Lett. 1986 Nov 24;208(2):451-4. doi: 10.1016/0014-5793(86)81067-5.
Protein kinase activity in isolated nuclei from rat liver was detected in situ after electrophoresis on SDS-polyacrylamide gel containing no exogenous protein substrate. After renaturation of polypeptides, the gel was incubated with [gamma-32P]ATP and divalent cations. Among five major protein kinase activities observed as radioactive bands by autoradiography, a protein kinase autophosphorylating on tyrosine (Mr 30,000) was identified and found to be localized in the nucleus, particularly in the nuclear matrix. The intensity of the activity band representing the level of the protein-tyrosine kinase in rat liver nuclei did not appreciably change during 3-24 h after partial hepatectomy.
在不含外源蛋白质底物的SDS-聚丙烯酰胺凝胶上进行电泳后，原位检测大鼠肝脏分离细胞核中的蛋白激酶活性。多肽复性后，将凝胶与[γ-32P]ATP和二价阳离子一起孵育。通过放射自显影观察到的五条主要蛋白激酶活性带中，鉴定出一种能在酪氨酸上进行自身磷酸化的蛋白激酶（分子量30,000），并发现其定位于细胞核中，尤其是核基质中。代表大鼠肝细胞核中蛋白酪氨酸激酶水平的活性带强度在部分肝切除术后3 - 24小时内没有明显变化。
