title: Changes in the mitochondrial phosphoproteome during mammalian hibernation.
abstract: Mammalian hibernation involves periods of substantial suppression of metabolic rate (torpor) allowing energy conservation during winter. In thirteen-lined ground squirrels (Ictidomys tridecemlineatus), suppression of liver mitochondrial respiration during entrance into torpor occurs rapidly (within 2 h) before core body temperature falls below 30°C, whereas reversal of this suppression occurs slowly during arousal from torpor. We hypothesized that this pattern of rapid suppression in entrance and slow reversal during arousal was related to changes in the phosphorylation state of mitochondrial enzymes during torpor catalyzed by temperature-dependent kinases and phosphatases. We compared mitochondrial protein phosphorylation among hibernation metabolic states using immunoblot analyses and assessed how phosphorylation related to mitochondrial respiration rates. No proteins showed torpor-specific changes in phosphorylation, nor did phosphorylation state correlate with mitochondrial respiration. However, several proteins showed seasonal (summer vs. winter) differences in phosphorylation of threonine or serine residues. Using matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry, we identified three of these proteins: F1-ATPase α-chain, long chain-specific acyl-CoA dehydrogenase, and ornithine transcarbamylase. Therefore, we conclude that protein phosphorylation is likely a mechanism involved in bringing about seasonal changes in mitochondrial metabolism in hibernating ground squirrels, but it seems unlikely to play any role in acute suppression of mitochondrial metabolism during torpor.
authors.affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada. dchungch@zoology.ubc.ca
authors.forename: Dillon J
authors.initials: DJ
authors.lastname: Chung
authors.forename: Beata
authors.initials: B
authors.lastname: Szyszka
authors.forename: Jason C L
authors.initials: JC
authors.lastname: Brown
authors.forename: Norman P A
authors.initials: NP
authors.lastname: Hüner
authors.forename: James F
authors.initials: JF
authors.lastname: Staples
mesh_terms: D000818:Animals; D001831:Body Temperature; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006605:Hibernation; D008297:Male; D008322:Mammals / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism*; D010101:Oxygen Consumption; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012589:Sciuridae / Q000378:metabolism; D012621:Seasons; D012694:Serine / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013912:Threonine / Q000378:metabolism
chemical_list: D024101:Mitochondrial Proteins; D010750:Phosphoproteins; D020543:Proteome; D013912:Threonine; D012694:Serine
keywords: mitochondrial respiration; posttranslational modification; protein phosphorylation; thirteen-lined ground squirrel