Dickson R B, Willingham M C, Pastan I
J Cell Biol. 1981 Apr;89(1):29-34. doi: 10.1083/jcb.89.1.29.
alpha 2-Macroglobulin (alpha 2 M) was adsorbed to colloidal gold and used as a new tool in the study of receptor-mediated endocytosis. alpha 2 M-gold is easy to prepare and is clearly visualized at the electron microscope level. When cells were incubated with alpha 2 M-gold at 0 degrees C, gold was visualized both diffusely over the cell surface and concentrated in coated pits. After cells to which alpha 2 M-gold had been bound at 0 degrees C were warmed, the gold was rapidly internalized into uncoated vesicles, previously termed receptosomes. After 30 min of incubation or longer, gold was found in small lysosomes and, later, in large lysosomes and very small vesicles in the region of the Golgi complex. This pattern of localization is similar to that previously described, using peroxidase-labeled anti-alpha 2 M antibodies. By incubating cells with both alpha 2 M-gold and vesicular stomatitis virus (VSV), we studied the internalization of these two markers simultaneously. VSV and alpha 2 M-gold rapidly clustered in the same coated pits and were internalized in the same receptosomes. Proteins and hormones adsorbed to gold may be useful in the study of receptor-mediated endocytosis.
α2-巨球蛋白(α2M)被吸附到胶体金上,并用作研究受体介导的内吞作用的新工具。α2M-金易于制备,并且在电子显微镜水平下清晰可见。当细胞在0℃下与α2M-金孵育时,金在细胞表面呈弥散状可见,并集中在被膜小窝中。在0℃下已结合α2M-金的细胞升温后,金迅速内化到无被小泡中,这些小泡以前称为受体小体。孵育30分钟或更长时间后,在小溶酶体中发现了金,随后在高尔基体区域的大溶酶体和非常小的小泡中也发现了金。这种定位模式与先前使用过氧化物酶标记的抗α2M抗体所描述的模式相似。通过将细胞与α2M-金和水泡性口炎病毒(VSV)一起孵育,我们同时研究了这两种标记物的内化过程。VSV和α2M-金迅速聚集在相同的被膜小窝中,并在相同的受体小体中内化。吸附到金上的蛋白质和激素可能在受体介导的内吞作用研究中有用。