MOLECULAR BIOLOGY FOR SYNTHESIS OF GIANT PIGMENT GRANULES IN CONGENITAR PIGMENTARY DISEASES

先天性色素疾病中巨型色素颗粒合成的分子生物学

• 批准号: 12470179
• 负责人: JIMBOW Kowichi
• 金额: $ 9.15万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470179/
• 关键词: melanosome; tyrosinase; calnexin; giant granules; pigmentation; melanin; メラニン生合成; 分子シャペロン; 小胞輸送; チロジナーゼ

This study examines the target signals for the vesicular transport of tyrosinase and its related proteins (TRP-1,2) from Golgi (TGN) to melanosomes, in the hope to identify the biological and molecular mechanism involved in the synthesis of giant pigment granules in congenital pigmentary diseases. Specifically this study focused on characterization of the biological role of a low molecular weight GTP-binding protein, Rab7. To investigate the requirement of Rab7-containing compartments for vesicular transport of tyrosinase family proteins, we expressed tyrosinase and TRPs by recombinant adenovirus and analyzed their localization in human amelanotic melanoma cells in the presence or absence of a dominant-negative mutant of Rab7 (Rab7N125I). Co-infection (Ad-HT) and TRP-1 (Ad-TRP-1) resulted in the enhancement of tyrosinase activity and melanin production compared to a single infection of Ad-HT. In the Ad-HT-infected cells many of the newly synthesized tyrosinase proteins were colocalized in lysosomal Igp85-positive granules of the entire cytoplasm, whereas in the presence of Rab7N125I the colocalization tyrosinase and Igp85 proteins was decreased markedly in the distal area of the cytoplasm. In the Ad-TRP-1-infected cells, TRP-1 was detected throughout the cytoplasm, but not colocalized in prelysosomal (early endosomal) EEA-1 granules. In the presence of Rab7N125I, however, TRP-1 was retained in the EEA-1-positive granules.We are in the process of analyzing the functional domains of TRP-1 and tyrosinase in this vesicular transport of the melanosome biosynthesis.

本研究旨在探讨酪氨酸酶及其相关蛋白（TRP-1，2）从高尔基体（Golgi，TGN）囊泡转运至黑素体的靶信号，以期明确先天性色素性疾病中巨大色素颗粒合成的生物学和分子机制。具体来说，这项研究集中在表征的生物作用的低分子量的GTP结合蛋白，Rab 7。为了研究酪氨酸酶家族蛋白的囊泡运输的Rab 7-含有车厢的要求，我们表达酪氨酸酶和TRP的重组腺病毒和分析其在人无色素性黑色素瘤细胞中的定位在Rab 7（Rab 7 N125 I）的显性阴性突变体的存在或不存在。联合感染（Ad-HT）和TRP-1（Ad-TRP-1）导致酪氨酸酶活性和黑色素生成比单独感染Ad-HT增强。在Ad-HT感染的细胞中，许多新合成的酪氨酸酶蛋白共定位于整个细胞质的溶酶体Igp 85阳性颗粒中，而在Rab 7 N125 I的存在下，共定位酪氨酸酶和Igp 85蛋白在细胞质的远端区域显着降低。在Ad-TRP-1感染的细胞中，TRP-1在整个细胞质中检测到，但不共定位于前溶酶体（早期内体）EEA-1颗粒。然而，在Rab 7 N125 I的存在下，TRP-1被保留在EEA-1阳性颗粒中。我们正在分析TRP-1和酪氨酸酶在黑素体生物合成的这种囊泡运输中的功能结构域。

项目成果

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Hirosaki K et al.: "Tyrosinase and tyrosinase related protein-1 (TRP-1) require Rab-7 for their intracellular transport"J Invest Dermatol.. 119. 475-480 (2002)

Hirosaki K 等人：“酪氨酸酶和酪氨酸酶相关蛋白 1 (TRP-1) 需要 Rab-7 进行细胞内运输”J Invest Dermatol.. 119. 475-480 (2002)

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Matsumoto Y 等：“从海胆磺基喹诺酮二酰基甘油中推导出的合成磺酰脂的免疫抑制作用”74・2（2002）。

Matsumoto Y et al.: "An immunosuppressive effect by synthetic sulfonolipids deduced from sulfonoquinovosly diacyiglycerols of sea urchin"Transplantation. 74・2. 261-267 (2002)

Matsumoto Y 等：“从海胆磺基喹诺酮二酰基甘油中推导出的合成磺酰脂的免疫抑制作用”74・2（2002）。

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Shinoda K 等人：“黑素体相关单克隆抗体 J1 识别黑素体和溶酶体生物发生中常见的前溶酶体腔膜”Cell Struct Fune。