气候变化领域集成服务门户(CCPortal): Dual-process brain mitochondria isolation preserves function and clarifies protein composition

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DOI	10.1073/pnas.2019046118
	Dual-process brain mitochondria isolation preserves function and clarifies protein composition
	Noterman M.F.; Chaubey K.; Lin-Rahardja K.; Rajadhyaksha A.M.; Pieper A.A.; Taylor E.B.
发表日期	2021
ISSN	00278424
卷号	118 期号:11
英文摘要	The brain requires continuously high energy production to maintain ion gradients and normal function. Mitochondria critically undergird brain energetics, and mitochondrial abnormalities feature prominently in neuropsychiatric disease. However, many unique aspects of brain mitochondria composition and function are poorly understood. Developing improved neuroprotective therapeutics thus requires more comprehensively understanding brain mitochondria, including accurately delineating protein composition and channel-transporter functional networks. However, obtaining pure mitochondria from the brain is especially challenging due to its distinctive lipid and cell structure properties. As a result, conflicting reports on protein localization to brain mitochondria abound. Here we illustrate this problem with the neuropsychiatric disease-associated L-type calcium channel Cav1.2α1 subunit previously observed in crude mitochondria. We applied a dual-process approach to obtain functionally intact versus compositionally pure brain mitochondria. One branch utilizes discontinuous density gradient centrifugation to isolate semipure mitochondria suitable for functional assays but unsuitable for protein localization because of endoplasmic reticulum (ER) contamination. The other branch utilizes self-forming density gradient ultracentrifugation to remove ER and yield ultrapure mitochondria that are suitable for investigating protein localization but functionally compromised. Through this process, we evaluated brain mitochondria protein content and observed the absence of Cav1.2α1 and other previously reported mitochondrial proteins, including the NMDA receptor, ryanodine receptor 1, monocarboxylate transporter 1, excitatory amino acid transporter 1, and glyceraldehyde 3-phosphate dehydrogenase. Conversely, we confirmed mitochondrial localization of several plasma membrane proteins previously reported to also localize to mitochondria. We expect this dual-process isolation procedure will enhance understanding of brain mitochondria in both health and disease. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Channel; Mitochondria; Neuropsychiatric disease; Solute carrier; Transporter
语种	英语
scopus关键词	calcium channel L type; excitatory amino acid transporter 1; functional group; glyceraldehyde 3 phosphate dehydrogenase; membrane protein; mitochondrial protein; monocarboxylate transporter 1; n methyl dextro aspartic acid receptor; ryanodine receptor 1; adult; animal experiment; Article; brain function; brain mitochondrion; cell structure; controlled study; density gradient centrifugation; endoplasmic reticulum; energy transfer; energy yield; female; glycosylation; human; immunoreactivity; male; mouse; nerve cell network; neuroprotection; nonhuman; priority journal; protein analysis; protein function; protein localization
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180249
作者单位	Interdisciplinary Graduate Program in Neuroscience, University of Iowa, Iowa City, IA 52242, United States; Harrington Discovery Institute, University Hospitals Cleveland Medical Center, Cleveland, OH 44106, United States; Department of Psychiatry, Case Western Reserve University, Cleveland, OH 44106, United States; Department of Systems Biology and Bioinformatics, Case Western Reserve University, Cleveland, OH 44106, United States; Weill Cornell Autism Research Program, Weill Cornell Medicine of Cornell University, New York, NY 10065, United States; Pediatric Neurology, Pediatrics, Weill Cornell Medicine of Cornell University, New York, NY 10065, United States; Geriatric Research Education and Clinical Centers, Louis Stokes Cleveland VA Medical Center, Cleveland, OH 44106, United States; Department of Neuroscience, School of Medicine, Case Western Reserve University, Cleveland, OH 44106, United States; Department of Molecular Physiology and Biophysics, Carver College of Medicine, University of Io...
推荐引用方式 GB/T 7714	Noterman M.F.,Chaubey K.,Lin-Rahardja K.,et al. Dual-process brain mitochondria isolation preserves function and clarifies protein composition[J],2021,118(11).
APA	Noterman M.F.,Chaubey K.,Lin-Rahardja K.,Rajadhyaksha A.M.,Pieper A.A.,&Taylor E.B..(2021).Dual-process brain mitochondria isolation preserves function and clarifies protein composition.Proceedings of the National Academy of Sciences of the United States of America,118(11).
MLA	Noterman M.F.,et al."Dual-process brain mitochondria isolation preserves function and clarifies protein composition".Proceedings of the National Academy of Sciences of the United States of America 118.11(2021).