JMSSJ On-line

JMSSJ On-line, Vol. 51 (2003) No. 5, pp. 509-515

Detection of Oxidized Proteins in Muscles of Diabetic Rats		>> Full Text PDF >> References
*Tadakazu MAEDA ^a), Masamichi OH-ISHI ^a), Takeshi UENO ^a), and Yoshio KODERA ^a)** ^a)Laboratory of Biomolecular Dynamics, Department ofPhysics, Kitasato University School of Science (1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan)*

Oxidative stress is implicated in a broad variety of chronic and acute diseases, including such age-related diseases as diabetes. To understand at the protein level cellular damage caused by the stress, we developed a proteomic method, in which protein carbonyls were derivatized with biotin hydrazide followed by two-dimensional gel electrophoresis with agarose gels in the first dimension. The method, being capable of analyzing high-molecular-mass proteins as large as myosin heavy chains (molecular mass: 200 kDa), was applied to detecting protein carbonyls in muscles of a diabetes model OLETF rat and a control LETO rat. A number of proteins, including mitochondrial ATP synthase β-chain, desmin, actin, and myosin, were found carbonylated. Our method would provide a means toward clarifying a comprehensive view of oxidative modifications of proteins during a long progression of age-related diseases and understanding the mechanism of the onset, progression, and complication of the diseases.

Key words: Protein carbonyls, Biotin hydrazide, Agarose 2-DE, LC-MS, Diabetes