Mitochondrial proteomics analysis of tumorigenic and metastatic breast cancer markers

Abstract

Mitochondria are key organelles in mammary cells responsible for several cellular functions including growth, division, and energy metabolism. In this study, mitochondrial proteins were enriched for proteomics analysis with the state-of-the-art two-dimensional differential gel electrophoresis and matrix-assistant laser desorption ionization-time-of-flight mass spectrometry strategy to compare and identify the mitochondrial protein profiling changes between three breast cell lines with different tumorigenicity and metastasis. The proteomics results demonstrate more than 1,500 protein features were resolved from the equal amount pooled from three purified mitochondrial proteins, and 125 differentially expressed spots were identified by their peptide finger print, in which, 33 identified proteins belonged to mitochondrial proteins. Eighteen out of these 33 identified mitochondrial proteins such as SCaMC-1 have not been reported in breast cancer research to our knowledge. Additionally, mitochondrial protein prohibitin has shown to be differentially distributed in mitochondria and in nucleus for normal breast cells and breast cancer cell lines, respectively. To sum up, our approach to identify the mitochondrial proteins in various stages of breast cancer progression and the identified proteins may be further evaluated as potential breast cancer markers in prognosis and therapy.

SCaMC-1 promotes cancer cell survival by desensitizing mitochondrial permeability transition via ATP/ADP-mediated matrix Ca2+ buffering

“…High levels of SCaMC-1 were previously found by screening approaches in colorectal carcinomas, 29 breast cancer 30 and in several cancer cell lines. 15,31 Our in silico studies confirmed and extended these findings to 67 different tumour types where SCaMC-1 expression significantly exceeded its expression in their normal tissue counterpart.…”

A Self-Sequestered Calmodulin-like Ca2+ Sensor of Mitochondrial SCaMC Carrier and Its Implication to Ca2+-Dependent ATP-Mg/Pi Transport

“…SCaMC is one of the two carriers responsible for transporting ATP. While AAC accounts for the bulk ADP/ATP recycling in the matrix by exporting newly synthesized ATP out of the matrix while importing ADP for continuous synthesis (Traba et al, 2009b), the transport activity of SCaMC is important for mitochondrial activities that depend on adenine nucleotides, such as gluconeogenesis, mitochondrial biogenesis and mitochondrial DNA maintenance (Amigo et al, 2013; Cavero et al, 2005; Chen et al, 2011; Kucejova et al, 2008; Llorente-Folch et al, 2013; Satrustegui et al, 2007). SCaMC is abundant, and to date, five major paralogs have been identified, SCaMC-1/SLC25A24, SCaMC-2/SLC25A25, SCaMC-3/SLC25A23 (Bassi et al, 2005; del Arco and Satrustegui, 2004; Fiermonte et al, 2004; Mashima et al, 2003), SCaMC-1L (Amigo et al, 2012) and SCaMC-3L (Traba et al, 2009a), most of which have several alternative spliced variants (del Arco and Satrustegui, 2004).…”

Proteomic analysis of UVB-induced protein expression- and redox-dependent changes in skin fibroblasts using lysine- and cysteine-labeling two-dimensional difference gel electrophoresis

“…Moreover, these protein visualization methods often have narrow linear dynamic ranges of detection, making them unsuitable for the analysis of biological samples where protein copy numbers vary enormously. A significant improvement in the ability to use gel-based methods for protein quantitation and detection was achieved with the introduction of 2D-difference gel electrophoresis (2D-DIGE), which enables co-detection of several samples on the same 2-DE gel, so avoiding gel-to-gel variation [16][17][18][19][20].…”