A molecular mechanism for copper transportation to tyrosinase that is assisted by a metallochaperone, caddie protein

铜转运至酪氨酸酶的分子机制，由金属伴侣、球童蛋白协助

• 批准号: 22550153
• 负责人: SUGIYAMA Masanori
• 金额: $ 3.08万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22550153/
• 关键词: チロシナーゼ; X 線結晶構造解析; 銅イオン; 金属シャペロン; メラニン生成酵素; 放線菌; タンパク質; ラマンスペクトル; 吸収スペクトル; X線結晶構造解析; チロシナーゼ; X 線結晶構造解析; 銅イオン; 金属シャペロン; メラニン生成酵素; 放線菌; タンパク質; ラマンスペクトル; 吸収スペクトル; X線結晶構造解析

The Cu(II)-soaked crystal structure of tyrosinase in a complex with a protein, designated “caddie”, which has previously determined by us, displays to possess two copper ions at the catalytic center. We observed two copper-binding sites in the caddie protein, speculating that the copper bound to caddie may be transported to the tyrosinase catalytic center. In the present study, at the 1.16 to 1.58 A resolution, we determined the crystal structures of tyrosinase complexed with a caddie prepared by altering the soaking time of the copper ion and the structures of tyrosinase complexed with a few mutants of a caddie which displays no or little ability to activate tyrosinase. Based on these structures, we propose a molecular mechanism to transport two copper ions to the tyrosinase catalytic center, which is assisted by a caddie as a metallochaperone.

在与我们以前已确定的蛋白质的蛋白质中，酪氨酸酶的铜（II）浸泡的晶体结构显示在催化中心中具有两个铜离子。我们观察到球童蛋白中的两个铜结合位点，猜测与球童结合的铜可能被运输到酪氨酸酶催化中心。在介绍研究中，在1.16至1.58 A分辨率下，我们通过改变铜离子的浸泡时间和与CADDY的几个突变体复合的酪氨酸酶结构来确定酪氨酸酶的晶体结构，该酪氨酸酶的浸泡时间和酪氨酸酶的结构没有表现出激活酪氨酸酶的能力或少量能力。基于这些结构，我们提出了一种分子机制，将两个铜离子传输到酪氨酸酶催化中心，该中心由球童作为金属伴侣的辅助。