Project description:Proteomic and Metabolomic analysis of exercise-induced sweat to evaluate analyte correlation with human performance parameters. Sweat was collected from participants on a treadmill at low, medium, and hard speed and incline. Sweat was analyzed by untagged HILIC LC-MS.
Project description:During exercise there is a site-specific increase in ROS within muscle required for the beneficial adaptive response by activation of specific signalling pathways. Peroxiredoxin 2 (Prdx2), an abundant cytoplasmic 2-Cys peroxidase, is required for the adaptive beneficial hormesis response to H2O2, myoblasts with knockdown of Prdx2 did not increase mitochondrial content and had disrupted myogenesis. Using a 5-day swimming protocol in C. elegans, previously been demonstrated to improve healthspan, we observed increased mitochondrial content, fitness, survival and longevity in wild type (N2) worms. However, prdx-2 mutant worms had decreased fitness, disrupted mitochondria, reduced survival and lifespan following exercise. Global proteomics following exercise revealed an increase in the abundance of proteins involved in fatty oxidation and ion transport, decrease in proteins involved in cuticle formation in N2 worms. In the prdx-2 mutants following exercise there was an increase in proteins related to the mTOR pathway and decrease in transsulfuration and microRNA biogenesis proteins. Relative quantification of the reversible oxidation state of individual Cysteine (Cys) residues revealed an increased oxidation of specific Cys residues following exercise in prdx-2 mutant. A number of regulatory Cys residues of metabolic, cytoskeletal and calcium handling proteins are reduced in N2 worms. A redox signalling relay whereby the oxidative equivalents from ROS are transferred from evolutionary conserved Prdxs to target proteins, would confer specificity on redox signalling required for the beneficial adaptations to exercise.
Project description:A single bout of exercise followed by intake of carbohydrates leads to glycogen supercompensation in the prior exercised muscle. The molecular mechanisms underlying this well-known phenomenon remain elusive. Here we report that a single bout of exercise induces marked activation of glycogen synthase (GS) and AMP-activated protein kinase (AMPK) for several days beyond normalized muscle glycogen content in man. Acute muscle specific deletion of AMPK activity in mouse muscle abrogated the ability for glycogen supercompensation, providing genetic evidence that AMPK serves as essential driver for glycogen supercompensation. Muscle proteomic analyses revealed elevated glucose uptake capacity in the prior exercised muscle while key proteins in fat oxidation and glycolysis largely remained unchanged. The temporal order of these sustained cellular alterations induced by a single bout of exercise provide a mechanism to offset the otherwise tight feedback inhibition of glycogen synthesis and glucose uptake by glycogen, ultimately leading to muscle glycogen supercompensation.
Project description:The pathophysiological effects of a number of metabolic and age-related disorders can be prevented to some extent by exercise and increased physical activity. However, the molecular mechanisms that contribute to the beneficial effects of muscle activity remain poorly explored. Availability of a fast, inexpensive, and genetically tractable model for muscle activity and exercise will allow rapid identification and characterization of the molecular mechanisms that mediate the beneficial effects of muscle activity. Here, we report the development and characterization of an optogenetically-inducible muscle contraction (OMC) model in Drosophila larvae that we used to model acute exercise-like physiological responses. To characterize muscle-specific transcriptional responses to acute exercise, we performed bulk mRNA-sequencing, revealing striking similarities between acute exercise-induced genes in flies and those previously identified in humans. Our larval muscle contraction model opens a path for rapid identification and characterization of exercise-induced factors.
Project description:Gene expression microarrays accompanying "Proteomic and genomic characterization of a yeast model for Ogden syndrome" by Doerfel et al 2016 in press at Yeast. Naa10 is a Na-terminal acetyltransferase that, in a complex with its auxiliary subunit Naa15, co-translationally acetylates the a-amino group of newly synthetized proteins as they emerge from the ribosome. Roughly 40-50% of the human proteome is acetylated by Naa10, rendering this an enzyme with one of the most broad substrate ranges known. Recently, we reported an X-linked disorder of infancy, Ogden syndrome, in two families harboring a c.109T>C (p.Ser37Pro) variant in NAA10. In the present study we performed in-depth characterization of a yeast model of Ogden syndrome. Stress tests and proteomic analyses suggest that the S37P mutation disrupts Naa10 function and reduces cellular fitness during heat shock, possibly due to dysregulation of chaperone expression and accumulation. Microarray and RNA-seq revealed a pseudo-diploid gene expression profile in DNaa10 cells, likely responsible for a mating defect. In conclusion, the data presented here further support the disruptive nature of the S37P/Ogden mutation and identify affected cellular processes potentially contributing to the severe phenotype seen in Ogden syndrome.
Project description:Fish species display huge differences in physical activity ranging from lethargy to migration of thousands of miles, making them an interesting model for human exercise. Here, we show a remarkable plasticity of zebrafish in response to exercise and induction of PGC1α (encoded by PPARGC1A), a dominant regulator of mitochondrial biogenesis. Forced expression of human PPARGC1A induces mitochondrial biogenesis, an exercise-like gene expression signature, and physical fitness comparable to wild-type animals trained in counter-current swim tunnels. We quantify a stoichiometric induction of the electron transport chain (ETC) in response to exercise or PGC1α expression, identified by a proteomic SWATH-MS workflow. Exercise or PGC1α expression induce the re-organization of the ETC into respiratory supercomplexes, and we show that ndufa4/ndufa4l, previously assigned to complex I, associates to free and supramolecular complex IV in vivo. Thus, zebrafish is a useful and experimentally tractable vertebrate model to study exercise biology, including ETC expression and assembly.
Project description:In this study, we undertook a global proteomics analysis of isolated PSII complexes, comparing protein profiles of HT3 (C-terminal hexahistidine tagged CP47 protein) to those of deltaPsbV HT3, deltaPsbP HT3, and deltaPsbQ HT3. The sensitivity of these techniques allowed for identification of not only stoichiometric components of active PSII complexes, but also for the identification of proteins that are transiently associated with PSII throughout its lifecycle, such as assembly, repair, or degradation partners. From the results, we identified an operon of unknown function which contains binding domains for photosynthetic cofactors and which we have named the cofactor integration operon (cio). Upon deletion of the operon, photosynthetic capacity is decreased, indicating a function in PSII-mediated activity. Keywords: proteomic, photosystem II, cyanobacteria
Project description:The mechanisms underlying exercise-induced effects in the skeletal muscle during cancer cachexia progression have not been fully described. Here, we tested the hypothesis that different exercise training protocols could attenuate metabolic impairment in a severe model of cancer cachexia. Moderate-intensity training (MIT) and high-intensity interval training (HIIT) improved running capacity and prolonged lifespan in tumor-bearing rats. HIIT also reduced oxidative stress and reestablished muscle contractile function. An unbiased proteomics screening revealed that COP9 signalosome complex subunit 2 (COPS2), also known as thyroid receptor interacting protein 15 (TRIP15) or ALIEN, is one of the most downregulated proteins at the early stage of cancer cachexia progression. HIIT restored COPS2/TRIP15/ALIEN protein expression to the control levels. Moreover, lung cancer patients with low endurance capacity had lower muscle COPS2/TRIP15/ALIEN protein content compared to age- and sex-matched control subjects. We further established an in vitro model of cancer-induced muscle wasting using tumor cells-conditioned media to explore the potential protective role of COPS2/TRIP15/ALIEN for myotubes homeostasis. This in vitro model indicate that tumor cells produce factors that directly affect myotube metabolism, but COPS2/TRIP15/ALIEN overexpression is not able to fully reestablish metabolic homeostasis and protein content in myotubes incubated with tumor cells-conditioned media. The current study provides new insight into the role of exercise training as a co-therapy for cancer cachexia and uncovers COPS2/TRIP15/ALIEN as a novel potential target for cancer cachexia.