...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Cytoskeleton >Positively charged residues within the MYO19 MyMOMA domain are essential for proper localization of MYO19 to the mitochondrial outer membrane
【24h】

Positively charged residues within the MYO19 MyMOMA domain are essential for proper localization of MYO19 to the mitochondrial outer membrane

机译：MYO19 MyMOMA域内带正电荷的残基对于MYO19正确定位到线粒体外膜至关重要

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Myosins are well characterized molecular motors essential for intracellular transport. MYO19 copurifies with mitochondria, and can be released from mitochondrial membranes by high pH buffer, suggesting that positively-charged residues participate in interactions between MYO19 and mitochondria. The MYO19-specific mitochondria outer membrane association (MyMOMA) domain contains approximately 150 amino acids with a pI approximately 9 and is sufficient for localization to the mitochondrial outer membrane. The minimal sequence and specific residues involved in mitochondrial binding have not been identified. To address this, we generated GFP-MyMOMA truncations, establishing the boundaries for truncations based on sequence homology. We identified an 83-amino acid minimal binding region enriched with basic residues (pI approximate to 10.5). We sequentially replaced basic residues in this region with alanine, identifying residues R882 and K883 as essential for mitochondrial localization. Constructs containing the RK882-883AA mutation primarily localized with the endoplasmic reticulum (ER). To determine if ER-associated mutant MyMOMA domain and mitochondria-associated wild type MyMOMA display differences in kinetics of membrane interaction, we paired FRAP analysis with permeabilization activated reduction in fluorescence (PARF) analysis. Mitochondria-bound and ER-bound MYO19 constructs displayed slow dissociation from their target membrane when assayed by PARF; both constructs displayed exchange within their respective organelle networks. However, ER-bound mutant MYO19 displayed more rapid exchange within the ER network than did mitochondria-bound MYO19. Taken together these data indicate that the MyMOMA domain contains strong membrane-binding activity, and membrane targeting is mediated by a specific, basic region of the MYO19 tail with slow dissociation kinetics appropriate for its role(s) in mitochondrial network dynamics. (c) 2016 Wiley Periodicals, Inc.
机译：肌球蛋白是细胞内转运必不可少的分子马达。 MYO19与线粒体共纯化，并可以通过高pH缓冲液从线粒体膜上释放，表明带正电荷的残基参与MYO19与线粒体之间的相互作用。 MYO19特异性线粒体外膜缔合（MyMOMA）域包含约150个氨基酸，pI约为9，足以定位于线粒体外膜。尚未确定参与线粒体结合的最小序列和特定残基。为了解决这个问题，我们生成了GFP-MyMOMA截短，并基于序列同源性建立了截短的边界。我们确定了一个83个氨基酸的最小结合区，富含碱性残基（pI约为10.5）。我们依次用丙氨酸替换了该区域中的基本残基，确定了残基R882和K883是线粒体定位所必需的。包含RK882-883AA突变的构建体主要位于内质网（ER）。若要确定ER相关突变MyMOMA域和线粒体相关野生型MyMOMA是否在膜相互作用动力学方面显示差异，我们将FRAP分析与透化激活的荧光还原（PARF）分析配对。当通过PARF检测时，线粒体结合和ER结合的MYO19构建体显示出从其靶膜缓慢解离。两种结构在其各自的细胞器网络内均显示出交换。但是，与线粒体结合的MYO19相比，与ER结合的突变体MYO19在ER网络中显示出更快的交换。这些数据加在一起表明MyMOMA结构域具有强大的膜结合活性，并且膜靶向作用是由MYO19尾巴的特定基本区域介导的，其慢解离动力学适合于其在线粒体网络动力学中的作用。 （c）2016年威利期刊有限公司

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号