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Thesis
Blue native 電気泳動法を用いたミトコンドリア呼吸鎖異常症の診断

本多　正和
埼玉医科大学　小児科部門

医学博士　甲第1041号　平成19年3月23日 （埼玉医科大学）

RBlue native polyacrylamide gel electrophoresis is a powerful tool for screening of mitochondrial respiratory chain disorders.
Masakazu Honda (Department of Pediatrics, School of Medicine, Saitama Medical University, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　Abstract:Mitochondrial respiratory chain disorders are not only the most common but the most clinically diverse group of inborn errors of metabolism. It is often difficult to establish an accurate diagnosis and then to determine the molecular basis of respiratory chain disorders. Blue native polyacrylamide gel electrophoresis (BN-PAGE) is a form of native electrophoresis for resolving membrane protein complexes. We used BN-PAGE combined either with in gel enzyme staining or with immunoblotting to analyze 20 patients with possible, probable or definite mitochondrial respiratory chain disorders as defined by the criteria of Bernier et al. A potential complex I abnormality was detected in 11 out of 20 patients, but no abnormalities were found either in the amounts or activities of other complexes. Complex I abnormalities were categorized into three groups based on amount and size of assembled complex I. Combined with conventional enzyme assay, 4 out of 11 patients having complex I abnormality in BN-PAGE analyses were diagnosed as definite complex I deficiency. Tissue specificity in three sisters with complex I deficiency was also identified. We conclude that BN-PAGE is a powerful tool both in the correct diagnosis and as a useful guide to future molecular analysis for respiratory chain disorders.
Keywords: Blue native polyacrylamide gel electrophoresis; Mitochondrial respiratory chain disorders; In gel enzyme staining; Immunoblotting; Tissue specificity