Proteomic and bioinformatic characterization of the biogenesis and function of melanosomes
Proteomic and bioinformatic characterization of the biogenesis and function of melanosomes
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DOI：
10.1021/pr060363j
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发表时间：
2006-11-03
影响因子：
4.4
通讯作者：
Hunt, Donald F.
中科院分区：
文献类型：
--
作者：
Chi, An;Valencia, Julio C.;Hunt, Donald F.
黑色素几乎负责人类所有可见的皮肤、头发和眼睛色素沉着，它是在称为黑素体的亚细胞细胞器中合成、沉积和分布的。存在的黑色素阻碍了对该细胞器蛋白质组成的全面测定。在这里，我们报告了一种去除黑色素的新方法，包括溶液内消化和固定化金属亲和层析（IMAC）。与凝胶内消化一起，该方法使我们能够通过串联质谱法表征不同发育阶段的黑素体蛋白质组。黑素体蛋白质组的比较分析和功能表征在所有阶段的黑素体中鉴定出与 1500 种蛋白质相似的蛋白质，在任何给定阶段中与 600 种蛋白质相似。这些蛋白质包括 16 个与小鼠皮毛颜色基因同源的蛋白质，以及许多与人类色素疾病相关的蛋白质。来自色素和非色素黑素细胞的黑素体共享大约 100 种蛋白质，定义了必需的黑素体蛋白质组。通过确认其细胞内定位而验证的蛋白质包括 PEDF（色素上皮衍生因子）和 SLC24A5（钠/钾/钙交换器 5，NCKX5）。黑素体和其他溶酶体相关细胞器之间的蛋白质共享表明了共同的进化起源。这项工作代表了溶酶体相关细胞器生物发生研究的模型。
Melanin, which is responsible for virtually all visible skin, hair, and eye pigmentation in humans, is synthesized, deposited, and distributed in subcellular organelles termed melanosomes. A comprehensive determination of the protein composition of this organelle has been obstructed by the melanin present. Here, we report a novel method of removing melanin that includes in-solution digestion and immobilized metal affinity chromatography ( IMAC). Together with in-gel digestion, this method has allowed us to characterize melanosome proteomes at various developmental stages by tandem mass spectrometry. Comparative profiling and functional characterization of the melanosome proteomes identified similar to 1500 proteins in melanosomes of all stages, with similar to 600 in any given stage. These proteins include 16 homologous to mouse coat color genes and many associated with human pigmentary diseases. Approximately 100 proteins shared by melanosomes from pigmented and nonpigmented melanocytes define the essential melanosome proteome. Proteins validated by confirming their intracellular localization include PEDF (pigment-epithelium derived factor) and SLC24A5 (sodium/potassium/calcium exchanger 5, NCKX5). The sharing of proteins between melanosomes and other lysosome-related organelles suggests a common evolutionary origin. This work represents a model for the study of the biogenesis of lysosome-related organelles.