Absolute quantitative profiling of the key metabolic pathways in slow and fast skeletal muscle
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Absolute quantitative profiling of the key metabolic pathways in slow and fast skeletal muscle. / Rakus, Dariusz; Gizak, Agnieszka; Deshmukh, Atul; Wiśniewski, Jacek R.
In: Journal of Proteome Research, Vol. 14, No. 3, 06.03.2015, p. 1400-11.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Absolute quantitative profiling of the key metabolic pathways in slow and fast skeletal muscle
AU - Rakus, Dariusz
AU - Gizak, Agnieszka
AU - Deshmukh, Atul
AU - Wiśniewski, Jacek R
PY - 2015/3/6
Y1 - 2015/3/6
N2 - Slow and fast skeletal muscles are composed of, respectively, mainly oxidative and glycolytic muscle fibers, which are the basic cellular motor units of the motility apparatus. They largely differ in excitability, contraction mechanism, and metabolism. Because of their pivotal role in body motion and homeostasis, the skeletal muscles have been extensively studied using biochemical and molecular biology approaches. Here we describe a simple analytical and computational approach to estimate titers of enzymes of basic metabolic pathways and proteins of the contractile machinery in the skeletal muscles. Proteomic analysis of mouse slow and fast muscles allowed estimation of the titers of enzymes involved in the carbohydrate, lipid, and energy metabolism. Notably, we observed that differences observed between the two muscle types occur simultaneously for all proteins involved in a specific process such as glycolysis, free fatty acid catabolism, Krebs cycle, or oxidative phosphorylation. These differences are in a good agreement with the well-established biochemical picture of the muscle types. We show a correlation between maximal activity and the enzyme titer, suggesting that change in enzyme concentration is a good proxy for its catalytic potential in vivo. As a consequence, proteomic profiling of enzyme titers can be used to monitor metabolic changes in cells. Additionally, quantitative data of structural proteins allowed studying muscle type specific cell architecture and its remodeling. The presented proteomic approach can be applied to study metabolism in any other tissue or cell line.
AB - Slow and fast skeletal muscles are composed of, respectively, mainly oxidative and glycolytic muscle fibers, which are the basic cellular motor units of the motility apparatus. They largely differ in excitability, contraction mechanism, and metabolism. Because of their pivotal role in body motion and homeostasis, the skeletal muscles have been extensively studied using biochemical and molecular biology approaches. Here we describe a simple analytical and computational approach to estimate titers of enzymes of basic metabolic pathways and proteins of the contractile machinery in the skeletal muscles. Proteomic analysis of mouse slow and fast muscles allowed estimation of the titers of enzymes involved in the carbohydrate, lipid, and energy metabolism. Notably, we observed that differences observed between the two muscle types occur simultaneously for all proteins involved in a specific process such as glycolysis, free fatty acid catabolism, Krebs cycle, or oxidative phosphorylation. These differences are in a good agreement with the well-established biochemical picture of the muscle types. We show a correlation between maximal activity and the enzyme titer, suggesting that change in enzyme concentration is a good proxy for its catalytic potential in vivo. As a consequence, proteomic profiling of enzyme titers can be used to monitor metabolic changes in cells. Additionally, quantitative data of structural proteins allowed studying muscle type specific cell architecture and its remodeling. The presented proteomic approach can be applied to study metabolism in any other tissue or cell line.
KW - Animals
KW - Chromatography, Liquid
KW - Citric Acid Cycle
KW - Fatty Acids
KW - Female
KW - Glycolysis
KW - Mice
KW - Mice, Inbred C57BL
KW - Muscle Fibers, Fast-Twitch
KW - Muscle Fibers, Slow-Twitch
KW - Oxidative Phosphorylation
KW - Pyruvate Dehydrogenase Complex
KW - Tandem Mass Spectrometry
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1021/pr5010357
DO - 10.1021/pr5010357
M3 - Journal article
C2 - 25597705
VL - 14
SP - 1400
EP - 1411
JO - Journal of Proteome Research
JF - Journal of Proteome Research
SN - 1535-3893
IS - 3
ER -
ID: 170597865