Control Mechanisms for Matching ATP Supply and Demand in Heart Mitochondria

心脏线粒体中 ATP 供需匹配的控制机制

• 批准号: 9348184
• 负责人: Steven Sollott
• 金额: $ 53.91万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9348184
• 关键词: ATP Synthesis Pathway; BCL-2 Protein; BCL2 gene; BH3 Domain; Bioenergetics; Calcium; Cardiac; Cardiac Myocytes; Cell Respiration; Cells; Consumption; Contracts; Coupled; Cytoprotection; Cytosol; Defect; Development; Dimensions; Elderly; Electric Stimulation; Electron Transport; Electrons; Elements; Energy Supply; Environment; Enzymes; Failure; Family; Frequencies; Heart; Heart Mitochondria; Heart failure; Homeostasis; Hydrolysis; Inner mitochondrial membrane; Ischemia; Lead; Link; Mechanics; Mitochondria; Modeling; Molecular; Myocardial; Oxidation-Reduction; Oxygen Consumption; Pathology; Persons; Phylogenetic Analysis; Physiological; Play; Population; Positioning Attribute; Potassium Channel; Potential Energy; Production; Protein Family; Proteins; Proton-Motive Force; Protons; Regulation; Reperfusion Injury; Respiration; Rest; Role; Running; Signal Transduction; Source; Specificity; Stress; Suspension substance; Suspensions; Time; Tissues; Trees; Weight; Work; Workload; effective therapy; meetings; member; metabolic rate; molecular recognition; novel; reaction rate; response

Failure to supply energy to match the body's demands limits the functional reserve capacity, and under certain periods of stress, such as ischemia, can lead to irreversible cell and tissue damage. This matching is critical in tissues with high and rapidly fluctuating metabolic rates such as the heart. Mitochondria are the main ATP suppliers to meet cellular demands. The fuel used by mitochondria is transported across the inner mitochondrial membrane to the matrix and produces a source of electrons whose redox-potential energy is, in turn, harnessed by the electron transport chain. The flux of electrons is reflected in oxygen consumption. The energy released from this electron flow is used to transport protons out of the matrix across the inner mitochondrial membrane forming a gradient whose proton-motive force drives ATP synthase to make ATP. This "upstream" regulation is known as the "push" mechanism. A complete description of the ATP synthase control mechanisms is still lacking. Two general mechanisms have been suggested to serve as key regulators: 1) ADP and Pi concentrations; ATP utilization/hydrolysis in the cytosol increases ADP and Pi fluxes to mitochondria and hence the amount of available substrates for ATP production increases; 2) Ca2+ concentration; ATP utilization/hydrolysis is coupled to changes in free cytosolic calcium and mitochondrial Ca2+, the latter controlling Ca2+-dependent activation of certain mitochondrial reaction-rate-determining enzymes taking part in ATP production. At high levels of energy demand the question arises whether parallel to the "push" mechanism signals acting on ATP synthase could also facilitate the electron transport chain redox flux, enhancing the efficiency of ATP production. This effect simulates an apparent additional "pull" on the upstream flux, which causes as a specific proportionate increase in respiration. Proof of such a "pull" mechanism regulated by Ca2+ and its target has not been demonstrated to-date. Cardiomyocytes contract in response to driven cyclic 'increases' in cytosolic Ca2+ in a response to electrically stimulation. As a consequence of the levels of contractile work, ATP is proportionately utilized by the contractile elements. Therefore, from the demand point of view Ca2+ is a direct effector that might be well positioned to play a role in the energy matching regulatory mechanisms. A correlation has also been shown between cytosolic and mitochondrial Ca2+. Ca2+ enters the mitochondria through the mitochondrial uniporter and is extruded by the mitochondrial Na+/Ca2+ exchanger. This results in mitochondrial Ca2+ accumulation in response to an increase in stimulation frequency or Ca2+ transient amplitude. Therefore, Ca2+ levels in the mitochondria reflect changes in both myocardial work and ATP consumption and, hence, the demand for ATP. It was shown that mitochondrial Ca2+ can activate the mitochondrial enzymes taking part in ATP production. Therefore, changes in mitochondrial Ca2+ during electrical stimulation are linked to changes in ATP supply and demand. We and others have shown that small changes in mitochondrial volume can regulate respiration and in turn energy production. It is also known that the Ca2+ environment may regulate mitochondrial volume in isolated mitochondrial suspension raising the question whether physiological changes in Ca2+ via increasing electrically stimulated Ca2+ cycling would act in this way. We found that while increasing electrically stimulated, physiological Ca2+ cycling does not detectibly change the 'diastolic' mitochondrial long- and short-axis dimensions (i.e, volume) shortly (2.5 min) after the transition from rest to low or higher workloads, it nevertheless caused an increase in cell respiration (and in turn facilitated energy production) in both conditions. These results were in contrast to that observed by others in the isolated mitochondria models. Additionally, we found that the mechanisms that control ATP supply from the hearts mitochondria consist of both 'push' and 'pull' mechanisms and that 'pull' mechanism directly targets ATP synthase. We identified that the 'pull' mechanism is controlled by mitochondrial Ca2+ and can be further facilitated by pharmacologically regulating mitochondrial volume. At low cardiac workload, the 'push' mechanism is sufficient to match ATP supply and demand, and the mitochondrial transmembrane ADP/Pi gradient is presumably sufficient to drive the 'push' and 'pull' mechanisms. However, under the same experimental conditions, pharmacological induction of a regulatory mitochondrial volume increase was found to facilitate mitochondrial Ca2+ entry responsible for further pushing respiration, whereas at higher workloads, mitochondrial Ca2+ entry did not require such facilitation, and in turn was sufficient and essential to drive both "push" and 'pull' effects on respiration. Moreover, pharmacologically-enhanced mitochondrial Ca2+ accumulation (without changing cytosolic Ca2+) was also found to push respiration. Facilitation of these 'push' and 'pull' mechanisms is being examined as a potential treatment to reverse signaling defects in matching ATP supply and demand, such as occurs in heart failure which afflicts millions of people, especially the elderly population. We discovered that mammalian ATP synthase, previously believed to be a machine running exclusively on H+, actually utilizes almost 4 K+ for every H+ to make ATP inside intact cellular mitochondria. Thus, ATP synthase is, for the first time, identified as a primary mitochondrial K+ uniporter, i.e., the primary way for K+ to enter mitochondria; furthermore, since this K+ entry is directly proportional to ATP synthesis and regulates matrix volume, this in turn serves the function of directing the matching of cellular energy utilization with its production. For the first time, we show that the chemo-mechanical efficiency of ATP synthase can be up-regulated, and that this occurs by certain members of the Bcl-2 family and by certain K+ channel openers acting via an intrinsic regulatory factor of ATP synthase, IF1, which we identified as itself a novel and previously unrecognized member of the Bcl-2 protein family. As a consequence of the foregoing, we discovered that ATP synthase is also a recruitable mitochondrial ATP-dependent K+ channel which serves critical functions in cell protection signaling that can limit the damage of ischemia-reperfusion injury. Thus, we discovered the molecular identity of two mitochondrial potassium channels, an entirely new function set for ATP synthase, and what is likely the primary mechanism by which mitochondrial function matches energy supply with demand for all cells in the body. We discovered that IF1 is a novel, highly conserved BH3-only member of the Bcl-2 protein family displaying, in addition to the BH3 linear sequence motif, a functional BH3-domain-like molecular recognition feature (MoRF) which enables the modulation of ATP synthase function. The phylogenetic tree shows that IF1s linear motif is most closely related to the BH3-only proteins (e.g. Bak, Bid, etc.). These findings will fundamentally change our understanding of the regulation of mitochondrial energy production and homeostasis. Because we now know the identity of the mitochondrial K+-uniporter to be the ATP synthase, and given its dominant permeation by K+ over H+ to make the daily equivalent of the bodys weight in ATP, the actual rate and volume of mitochondrial K+ flux cycling is huge (and not the previously believed trickle-leak).

不能提供与身体需求相匹配的能量限制了功能储备能力，在某些时期的压力下，如缺血，可能导致不可逆的细胞和组织损伤。这种匹配在代谢率高且波动迅速的组织（如心脏）中是至关重要的。线粒体是满足细胞需要的主要ATP供给者。线粒体使用的燃料通过线粒体内膜被输送到基质中，并产生电子源，这些电子源的氧化还原势能反过来又被电子传递链利用。电子的通量反映在氧气的消耗上。电子流释放的能量被用来将质子从基质中运输出来，穿过线粒体内膜，形成一个梯度，其质子动力驱动ATP合成酶制造ATP。这种“上游”监管被称为“推动”机制。对ATP合酶的控制机制尚缺乏完整的描述。两种一般机制被认为是关键的调节因子：1)ADP和Pi浓度；细胞质中ATP的利用/水解增加了线粒体中ADP和Pi的通量，因此可用于ATP生产的底物数量增加；2) Ca2+浓度；ATP的利用/水解与游离胞质钙和线粒体Ca2+的变化有关，后者控制某些线粒体反应速率决定酶的Ca2+依赖性激活，这些酶参与ATP的产生。在高水平的能量需求下，问题出现了，是否平行于“推动”机制的信号作用于ATP合酶也可以促进电子传递链的氧化还原通量，提高ATP生产的效率。这种效应模拟了上游通量上明显的额外“拉力”，这导致了呼吸的特定比例增加。这种由Ca2+及其靶标调节的“拉力”机制的证据迄今尚未得到证实。