MITOCHONDRIA FROZEN WITH TREHALOSE RETAIN BIOLOGICAL FUNCTIONS

用海藻糖冷冻线粒体保留生物功能

• 批准号: 7601060
• 负责人: DONALD DAVID NEWMEYER
• 金额: $ 1.09万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7601060
• 关键词: ATP Synthesis Pathway; Apoptosis; Bax protein; Biological Process; Buffers; Calcium; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Equilibrium; Freezing; Funding; Grant; In Vitro; Institution; Liver Mitochondria; Mannitol; Membrane Potentials; Mitochondria; Mitochondrial Proteins; Mus; Organelles; Outer Mitochondrial Membrane; Process; Protein Family; Protein Import; Proteins; Research; Research Personnel; Resources; Source; Standards of Weights and Measures; Sucrose; Swelling; Trehalose; United States National Institutes of Health; cytochrome c; pro-apoptotic protein; sugar

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In apoptosis, the Bcl-2 family proteins Bax and Bak disrupt the normal barrier function of the mitochondrial outer membrane (MOM), causing pro-apoptotic proteins such as cytochrome c, Omi and Smac to be released from the intermembrane space (IMS) into the cytoplasm. This process can be studied in vitro with freshly isolated mouse liver mitochondria maintained in a physiologically balanced buffer. The MOMs of these organelles remain intact, unless Bak is activated, e.g. by incubation with a BH3-only protein such as tBid. In contrast, the MOMs of mitochondria frozen/thawed in standard sucrose-mannitol buffers immediately become leaky, rendering them useless for apoptosis research. However, here we show that mitochondria frozen in buffer containing the sugar, trehalose, maintained their MOM integrity and responsiveness to BH3-only proteins, much like fresh mitochondria. Freezing in trehalose buffer also preserved biological functions such as ATP synthesis, calcium-induced swelling, transmembrane potential and mitochondrial protein import. The ability to store active mitochondria will facilitate research on apoptosis and other mitochondrial functions that rely on an intact MOM.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 在凋亡中，Bcl-2家族蛋白BAX和BAK破坏了线粒体外膜（MOM）的正常屏障功能，从而导致促凋亡蛋白（例如细胞色素C，OMI和SMAC）从膜间空间（IMS）中释放到细胞质中。可以通过在生理平衡的缓冲液中维持的新鲜分离的小鼠肝线粒体在体外研究此过程。除非激活Bak，否则这些细胞器的妈妈仍然完好无损，例如通过与仅BH3蛋白（例如TBID）孵育。相比之下，在标准的蔗糖甘露醇缓冲液中冷冻/解冻的线粒体妈妈立即变得漏水，使其无用，无法进行细胞凋亡研究。但是，我们在这里表明，线粒体在含糖，海藻糖的缓冲液中冷冻，保持了妈妈的正直和对仅BH3蛋白的反应性，就像新鲜的线粒体一样。海藻糖缓冲液中的冷冻还保留了生物学功能，例如ATP合成，钙诱导的肿胀，跨膜电位和线粒体蛋白的进口。存储主动线粒体的能力将促进依赖完整妈妈的细胞凋亡和其他线粒体功能的研究。