Published ahead of print on September 15, 2005, doi:10.1165/rcmb.2005-0114OC Am. J. Respir. Cell Mol. Biol., Volume 33, Number 6, December 2005, 636-642 A more recent version of this article appeared on December 1, 2005
Submitted on March 22, 2005 Expression and Carbonylation of Creative Kinase in the Quadriceps Femoris Muscles of COPD PatientsEsther Barreiro1,1 Respiratory Medicine, Muscle and Respiratory System Research Unit and Experimental Health Sciences Department, Hospital del mar, Barcelona, Catalonia, Spain; Critical Care and Respiratory Divisions, McGill University, Royal Victoria Hospital and Meakins-Christie Laboratories, Montreal, Quebec, Canada, 2 Respiratory Medicine, Muscle and Respiratory System Research Unit and Experimental Health Sciences Department, Hospital del mar, Barcelona, Catalonia, Spain, 3 Critical Care and Respiratory Divisions, McGill University, Royal Victoria Hospital and Meakins-Christie Laboratories, Montreal, Quebec, Canada * To whom correspondence should be addressed. E-mail: sabah.hussain{at}muhc.mcgill.ca.
Oxidative protein modification involving carbonylation has recently been identified as an important factor in skeletal muscle dysfunction in patients with chronic obstructive pulmonary disease (COPD). However, the exact identity of modified proteins inside limb muscles of COPD patients remains unknown. We used 2D electrophoresis, immunoblotting, and mass spectrometry to identify carbonylated proteins in the vastus lateralis muscle of 12 COPD patients and 6 control subjects. Both creatine kinase (CK) and carbonic anhydrase III (CAIII) were identified as being strongly carbonylated in this muscle in both groups of subjects. Total CK activity, CK protein expression, and the intensity of CK carbonylation were significantly greater in the muscles of COPD patients as compared to control subjects, whereas CAIII protein expression and intensity of carbonylation were similar in the two groups. In COPD patients, CK activity and protein expression correlated positively with FEV1 and VO2max, whereas the intensity of CK carbonylation correlated negatively with the same parameters. These results indicate that oxygen radicals selectively target CK and CAIII inside limb muscles of humans. The observation that the intensity of CK carbonylation correlates negatively with CK activity in limb muscles of COPD patients suggests that carbonylation may have a deleterious effect on CK activity and may contribute to impaired CK function in the limb muscles of these patients.
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