Araki K, Horikawa T, Chakraborty A K, Nakagawa K, Itoh H, Oka M, Funasaka Y, Pawelek J, Ichihashi M
Department of Dermatology, Kobe University School of Medicine, Japan.
Pigment Cell Res. 2000 Oct;13(5):332-6. doi: 10.1034/j.1600-0749.2000.130505.x.
Rab3A is a small guanosine triphosphate (GTP)-binding protein that has been recently implicated in intracellular vesicle transport and the secretion of neurotransmitters in neuronal cells. We demonstrate here that Rab3A is associated with melanosomes in pigment cells. Rab3A as well as Rabphilin3A, a putative target protein of Rab3A, were detected in the melanosome fraction, purified from B16 murine melanoma cells by sucrose density gradient ultracentrifugation. In contrast, Rab GDP dissociation inhibitor (GDI), a GDP/GTP exchange protein for Rab3A, was found in the cytosol fraction. Further studies using confocal laser scanning microscopy and immunoelectron microscopy revealed that immunoreactive Rab3A is localized in conjunction with the melanosomal membrane. These results suggest the possibility of involvement of Rab3A-Rabphilin3A complex, regulated by Rab GDI, in the intracellular transport of melanosomes in pigment cells.
Rab3A是一种小的鸟苷三磷酸(GTP)结合蛋白,最近被认为与细胞内囊泡运输以及神经元细胞中神经递质的分泌有关。我们在此证明Rab3A与色素细胞中的黑素小体相关。通过蔗糖密度梯度超速离心从B16小鼠黑色素瘤细胞中纯化得到的黑素小体组分中检测到了Rab3A以及Rab3A的一种假定靶蛋白Rabphilin3A。相反,Rab GDP解离抑制剂(GDI),一种Rab3A的GDP / GTP交换蛋白,存在于胞质溶胶组分中。使用共聚焦激光扫描显微镜和免疫电子显微镜的进一步研究表明,免疫反应性Rab3A与黑素小体膜结合定位。这些结果表明,由Rab GDI调节的Rab3A - Rabphilin3A复合物可能参与色素细胞中黑素小体的细胞内运输。