Protein composition and function of red and white skeletal muscle mitochondria
Protein composition and function of red and white skeletal muscle mitochondria
复制标题
DOI：
10.1152/ajpcell.00496.2010
复制
发表时间：
2011-06-01
影响因子：
5.5
通讯作者：
Balaban, Robert S.
中科院分区：
文献类型：
--
作者：
Glancy, Brian;Balaban, Robert S.
Glancy B，Balaban RS。红、白色骨骼肌线粒体的蛋白质组成与功能。美国生理学杂志细胞生理学300：C1280-C1290，2011年。首次发布于2011年2月2日; doi：10.1152/ajpcell.00496.2010。-红色和白色肌肉面临着非常不同的能量需求。然而，目前尚不清楚相对线粒体蛋白质表达是否在肌肉类型之间存在差异。采用Percoll梯度离心法分离红、白色猪骨骼肌线粒体。使用蓝色天然（BN）-PAGE、二维差异凝胶电泳（2D DIGE）、光谱和相对和绝对定量的同量异序标记（iTRAQ）测定蛋白质组成差异。使用BN-PAGE凝胶内活性测定比较复合物IV和V活性，并使用丙酮酸（P）+苹果酸（M）、谷氨酸（G）+ M和棕榈酰-肉毒碱（PC）+ M评估最大线粒体呼吸速率。在没有Percoll步骤的情况下，注意到白色线粒体的主要胞质蛋白污染。去除污染后，在红色和白色线粒体之间观察到非常少的蛋白质差异。BN-PAGE显示复合物I-V的亚基组成或复合物IV和V的活性没有差异。iTRAQ分析检测到358个线粒体蛋白，其中69个具有统计学差异。生理显著性可能较低：在25%差异下，检测到48种蛋白质;在50%差异下，检测到14种蛋白质;在100%差异下，检测到3种蛋白质。因此，任何变化都可以被认为是生理上适度的。一个不同的领域是脂肪代谢，其中四种β-氧化酶在红色线粒体中高出25%。这与红色线粒体中PC + M氧化速率比白色线粒体高40%相关，而P + M和G + M氧化无差异。这些数据表明，红色和白色肌纤维之间的代谢需求差异主要是由线粒体的数量匹配，而不是由线粒体本身的显着改变。
Glancy B, Balaban RS. Protein composition and function of red and white skeletal muscle mitochondria. Am J Physiol Cell Physiol 300: C1280-C1290, 2011. First published February 2, 2011; doi: 10.1152/ajpcell.00496.2010.-Red and white muscles are faced with very different energetic demands. However, it is unclear whether relative mitochondrial protein expression is different between muscle types. Mitochondria from red and white porcine skeletal muscle were isolated with a Percoll gradient. Differences in protein composition were determined using blue native (BN)-PAGE, two-dimensional differential in gel electrophoresis (2D DIGE), optical spectroscopy, and isobaric tag for relative and absolute quantitation (iTRAQ). Complex IV and V activities were compared using BN-PAGE in-gel activity assays, and maximal mitochondrial respiration rates were assessed using pyruvate (P) + malate (M), glutamate (G) + M, and palmitoyl-carnitine (PC) + M. Without the Percoll step, major cytosolic protein contamination was noted for white mitochondria. Upon removal of contamination, very few protein differences were observed between red and white mitochondria. BN-PAGE showed no differences in the subunit composition of Complexes I-V or the activities of Complexes IV and V. iTRAQ analysis detected 358 mitochondrial proteins, 69 statistically different. Physiological significance may be lower: at a 25% difference, 48 proteins were detected; at 50%, 14 proteins were detected; and 3 proteins were detected at a 100%. Thus any changes could be argued to be physiologically modest. One area of difference was fat metabolism where four beta-oxidation enzymes were similar to 25% higher in red mitochondria. This was correlated with a 40% higher rate of PC + M oxidation in red mitochondria compared with white mitochondria with no differences in P + M and G + M oxidation. These data suggest that metabolic demand differences between red and white muscle fibers are primarily matched by the number of mitochondria and not by significant alterations in the mitochondria themselves.