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Molecular and cellular biochemistry

Calreticulin is localized at mitochondria of rat cardiomyocytes and affected by furazolidone.


PMID 25087122

Abstract

Calreticulin (CRT) is a calcium-buffering protein which is predominantly located in endoplasmic reticulum. In the previous mitochondria proteome analysis, we accidentally found that CRT may be also localized at myocardial mitochondria and was upregulated in a rat model of furazolidone-induced dilated cardiomyopathy. To our knowledge, there has not yet been any report of its presence in mitochondria of any cell types. The present study aimed to determine whether CRT was located at the mitochondria of rat cardiomyocytes and whether the mitochondrial CRT was affected by furazolidone. Mitochondrial preparations were isolated from primary cultured neonatal rat cardiomyocytes and purified by differential centrifugation. The purity of mitochondria was assessed by the reduction or elimination of the immunoreactivities of markers for cytosol, nucleus, sarcolemma, and endoplasmic reticulum. Western blot analysis demonstrated the presence of CRT in purified mitochondria of rat cardiomyocytes. The distribution of CRT to mitochondria was further confirmed by immuno-electron microscopy, flow cytometry, and laser scanning confocal microscopy (double staining with MitoTracker Red and CRT-Alexa Fluor 488). Western blot analysis also demonstrated that the mitochondrial content of CRT was significantly enhanced by furazolidone treatment by 2.73xa0±xa00.13 fold (Pxa0

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