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Oxidation and structural perturbation of redox-sensitive enzymes in injured skeletal muscle
Authors:Pierce Anson P  de Waal Eric  McManus Linda M  Shireman Paula K  Chaudhuri Asish R
Institution:

aSam and Ann Barshop Institute for Longevity and Aging Studies, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA

bDepartment of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA

cDepartment of Biochemistry, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA

dDepartment of Pathology, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA

eDepartment of Periodontics, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA

fDepartment of Surgery, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA

gDepartment of Medicine, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA

hThe Geriatric Research Education and Clinical Center, South Texas Veterans Health Care System, San Antonio, TX 78229, USA

Abstract:Molecular events that control skeletal muscle injury and regeneration are poorly understood. However, inflammation associated with oxidative stress is considered a key player in modulating this process. To understand the consequences of oxidative stress associated with muscle injury, inflammation, and regeneration, hind-limb muscles of C57Bl/6J mice were studied after injection of cardiotoxin (CT). Within 1 day post-CT injection, polymorphonuclear neutrophilic leukocyte accumulation was extensive. Compared to baseline, tissue myeloperoxidase (MPO) activity was elevated eight- and fivefold at 1 and 7 days post-CT, respectively. Ubiquitinylated protein was elevated 1 day postinjury and returned to baseline by 21 days. Cysteine residues of creatine kinase (CK) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were irreversibly oxidized within 1 day post-CT injection and were associated with protein conformational changes that fully recovered after 21 days. Importantly, protein structural alterations occurred in conjunction with significant decreases in CK activity at 1, 3, and 7 days post-CT injury. Interestingly, elevations in tissue MPO activity paralleled the time course of conformational changes in CK and GAPDH. In combination, these results demonstrate that muscle proteins in vivo are structurally and functionally altered via the generation of reactive oxygen species produced during inflammatory events after muscle injury and preceding muscle regeneration.
Keywords:Creatine kinase  Glyceraldehyde-3-phosphate dehydrogenase  Cardiotoxin  BisANS  Skeletal muscle injury  Free radicals
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