Basrur Venkatesha, Yang Feng, Kushimoto Tsuneto, Higashimoto Youichiro, Yasumoto Ken-ichi, Valencia Julio, Muller Jacqueline, Vieira Wilfred D, Watabe Hidenori, Shabanowitz Jeffrey, Hearing Vincent J, Hunt Donald F, Appella Ettore
Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
J Proteome Res. 2003 Jan-Feb;2(1):69-79. doi: 10.1021/pr025562r.
Melanin is a heterogeneous biopolymer produced only by specific cells termed melanocytes, which synthesize and deposit the pigment in specialized membrane-bound organelles known as melanosomes. Although melanosomes have been suspected of being closely related to lysosomes and platelets, the total number of melanosomal proteins is still unknown. Thus far, six melanosome-specific proteins have been identified, and the challenge is to characterize the complete proteome of the melanosome to further understand its mechanism of biogenesis. In this report, we used mass spectrometry and subcellular fractionation to identify protein components of early melanosomes. Using this approach, we have identified all 6 of the known melanosome-specific proteins, 56 proteins that are shared with other organelles, and confirmed the presence of 6 novel melanosomal proteins using western blotting and by immunohistochemistry.
黑色素是一种仅由特定细胞即黑素细胞产生的异质生物聚合物,黑素细胞在称为黑素小体的特殊膜结合细胞器中合成并沉积色素。尽管黑素小体一直被怀疑与溶酶体和血小板密切相关,但黑素小体蛋白质的总数仍然未知。到目前为止,已经鉴定出六种黑素小体特异性蛋白质,而挑战在于表征黑素小体的完整蛋白质组以进一步了解其生物发生机制。在本报告中,我们使用质谱和亚细胞分级分离来鉴定早期黑素小体的蛋白质成分。通过这种方法,我们已经鉴定出所有6种已知的黑素小体特异性蛋白质、56种与其他细胞器共有的蛋白质,并通过蛋白质印迹法和免疫组织化学法证实了6种新的黑素小体蛋白质的存在。