(39) ALTERATION IN MITOCHONDRIAL PROTEINS IN MOUSE HEARTS MISSING CREATINE KINASE USING DIFFERENCE GEL ELECTROPHORESIS (DIGE)

Florence Kernec, Mustafa Ünlü, Wladimir Labeikovsky, Jonathan S. Minden, Alan Koretsky
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA

Difference Gel Electrophoresis (DIGE) is a new approach to detect proteomic changes [1]. This method has the unique advantage of requiring only a single gel to detect differences between two different fluorescent dyes prior to electrophoresis. Heart mitochondria have been isolated from sex and age matched C57B1/6, 129 S/v and mice missing both M- and mitochondrial forms of creatine kinase (mixed genetic C57B1/6x 129/Sv background) mice [2]. The purity of the isolated mitochondria preparation has been checked using electron microscopy. Our initial results using DIGE on isolated mitochondria show very consistent protein profiles between any two mitochondrial samples. Furthermore, no protein differences were detected between mitochondria isolated from two different mice of the same strain. Interestingly, when either C57B1/6, 129/Sv and DKO-CK are compared to each other, some distinct protein differences can be observed. Identification of these proteins by mass spectrometry is currently in progress in order to determine the changes in mitochondrial proteins caused by the elimination of muscle specific CK isoforms.

[1] Ünlü M et al. (1997) Electrophoresis 11:2072-2077. [2] Steeghs K et al. (1997) Cell 1:93-103.