title: Analysis of creatine kinase activity with evaluation of protein expression under the effect of heat and hydrogen peroxide.
abstract: Protein oxidation has detrimental effects on the brain functioning, which involves inhibition of the crucial enzyme, brain type creatine kinase (CKBB), responsible for the CK/phosphocreatine shuttle system. Here we demonstrate a susceptibility of CKBB to several ordinary stressors. In our study enzymatic activity of purified recombinant brain-type creatine kinase was evaluated. We assayed 30 nMconcentration of CKBB under normal and stress conditions. In the direction of phosphocreatine formation hydrogen peroxide and heat treatments altered CKBB activity down to 26 and 14%, respectively. Also, examination of immunoblotted membrane patterns by SDS-PAGE electrophoresis and western blot analysis showed a decrease in expression levels of intrinsic CKBB enzyme in HeLa andA549 cells. Hence, our results clearly show that cytosolic CKBB is extremely sensitive to oxidative stress and heat induced inactivation. Therefore, due to its susceptibility, this enzyme may be defined as a potential target in brain damage.
authors.forename: A D
authors.initials: AD
authors.lastname: Rakhmetov
authors.forename: Lee Sang
authors.initials: LS
authors.lastname: Pil
authors.forename: L I
authors.initials: LI
authors.lastname: Ostapchenko
authors.forename: Chae Ho
authors.initials: CH
authors.lastname: Zoon
mesh_terms: D045744:Cell Line, Tumor; D052276:Creatine Kinase, BB Form / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003600:Cytosol / Q000378:metabolism; D015870:Gene Expression; D006367:HeLa Cells; D006358:Hot Temperature; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry*; D018384:Oxidative Stress; D010725:Phosphocreatine / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014162:Transfection
chemical_list: D011993:Recombinant Fusion Proteins; D010725:Phosphocreatine; D006861:Hydrogen Peroxide; D052276:Creatine Kinase, BB Form