Price C M, Pettijohn D E
Exp Cell Res. 1986 Oct;166(2):295-311. doi: 10.1016/0014-4827(86)90478-7.
Monoclonal antibodies and human autoimmune sera specific for the nuclear mitotic apparatus protein (NuMA protein) were applied to study the structure of this protein and its intracellular distribution. The NuMA protein was purified using immuno-affinity columns. Studies on this large (250 kD) nuclear protein indicated that it is a highly asymmetric phosphoprotein. It is present in all mammalian cells examined and in those of some non-mammals. Immunofluorescence studies on fixed cells demonstrated that its intracellular distribution is essentially the same in all species at all stages of the cell cycle. Immunoblot (western blot) analysis showed that the size of the NuMA protein varies slightly in different species. At the onset of mitosis the NuMA protein redistributes from the nucleus to two centrosomal structures that later will become part of the mitotic spindle pole. This occurs at the time of nuclear breakdown and eventually leads to an accumulation of the NuMA protein at the polar region of the mitotic spindle. After anaphase the protein redistributes from the spindle polar region into the reforming nucleus and concentrates initially at the site where nuclear lamins and perichomatin have been reported to assemble. Living cells microinjected with fluorescent anti-NuMA antibodies were studied to examine parameters that effect the redistribution of the NuMA protein in vivo. These experiments indicate that microtubule assembly is essential for the NuMA protein to accumulate in the polar region.
应用针对核有丝分裂器蛋白（NuMA蛋白）的单克隆抗体和人类自身免疫血清来研究该蛋白的结构及其细胞内分布。使用免疫亲和柱纯化NuMA蛋白。对这种大型（250 kD）核蛋白的研究表明，它是一种高度不对称的磷蛋白。在所检测的所有哺乳动物细胞以及一些非哺乳动物细胞中均有存在。对固定细胞的免疫荧光研究表明，在细胞周期的所有阶段，其在所有物种中的细胞内分布基本相同。免疫印迹（western blot）分析表明，NuMA蛋白的大小在不同物种中略有差异。在有丝分裂开始时，NuMA蛋白从细胞核重新分布到两个中心体结构，这些结构随后将成为有丝分裂纺锤体极的一部分。这发生在核解体时，并最终导致NuMA蛋白在有丝分裂纺锤体的极区积累。后期之后，该蛋白从纺锤体极区重新分布到重新形成的细胞核中，并最初集中在据报道核纤层蛋白和周染色质组装的部位。对注射了荧光抗NuMA抗体的活细胞进行研究，以检查影响NuMA蛋白在体内重新分布的参数。这些实验表明，微管组装对于NuMA蛋白在极区积累至关重要。
