Ni-Komatsu Li, Orlow Seth J
The Ronald O. Perelman Department of Dermatology and Cell Biology, New York University School of Medicine, Dermatology Room H-100, NYU School of Medicine, 560 First Avenue, New York, NY 10016, USA.
Exp Eye Res. 2006 Mar;82(3):519-28. doi: 10.1016/j.exer.2005.08.013. Epub 2005 Sep 30.
The processing and trafficking of tyrosinase, a melanosomal protein essential for pigmentation, was investigated in a human epithelial 293 cell line that stably expresses the protein. The effects of the pink-eyed dilution (p) gene product, in which mutations result in oculocutaneous albinism type 2 (OCA2), on the processing and trafficking of tyrosinase in this cell line were studied. The majority of tyrosinase was retained in the endoplasmic reticulum-Golgi intermediate compartment and the early Golgi compartment in the 293 cells expressing the protein. Coexpression of p could partially correct the mistrafficking of tyrosinase in 293 cells. Tyrosinase was targeted to the late endosomal and lysosomal compartments after treatment of the cells with compounds that correct the tyrosinase mistrafficking in albino melanocytes, most likely through altering intracellular pH, while the substrate tyrosine had no effect on the processing of tyrosinase. Remarkably, this heterologous expression system recapitulates the defective processing and mistrafficking of tyrosinase observed in OCA2 albino melanocytes and certain amelanotic melanoma cells. Coexpression of other melanosomal proteins in this heterologous system may further aid our understanding of the details of normal and pathologic processing of melanosomal proteins.
在稳定表达该蛋白的人上皮293细胞系中,对酪氨酸酶(一种色素沉着所必需的黑素体蛋白)的加工和运输进行了研究。研究了粉红眼稀释(p)基因产物(其突变导致2型眼皮肤白化病(OCA2))对该细胞系中酪氨酸酶加工和运输的影响。在表达该蛋白的293细胞中,大部分酪氨酸酶保留在内质网-高尔基体中间区室和早期高尔基体区室。p的共表达可部分纠正293细胞中酪氨酸酶的运输错误。在用可纠正白化病黑素细胞中酪氨酸酶运输错误的化合物处理细胞后,酪氨酸酶靶向晚期内体和溶酶体区室,最有可能是通过改变细胞内pH值,而底物酪氨酸对酪氨酸酶的加工没有影响。值得注意的是,这种异源表达系统概括了在OCA2白化病黑素细胞和某些无色素性黑色素瘤细胞中观察到的酪氨酸酶加工缺陷和运输错误。在这种异源系统中共表达其他黑素体蛋白可能会进一步帮助我们了解黑素体蛋白正常和病理加工的细节。