Proteomic study to determination of relative expression of VDAC isoforms and characterization of hexokinase binding sites in rat, bovine and avian brain mitochondria / Estudo proteômico para determinação da expressão relativa das isoformas de VDAC e caracterização dos sítios de ligação da hexoquinase em mitocôndrias cerebrais de rato, boi e ave

AUTOR(ES)

Mirele Daiana Poleti

DATA DE PUBLICAÇÃO

2008

RESUMO

The voltage dependent anion selective channels (VDACs) are a group of proteins first identified in the mitochondrial outer membrane that are able to form hydrophilic pore structures in membranes. VDAC are known to play an essential role in cellular metabolism and in the early stages of apoptosis. In mammals, three VDACs isoforms (VDAC1, 2, 3) have been identified. A proteomic approach, consisting of two dimensional electrophoresis, followed by western blotting with anti-VDAC 1, anti-VDAC 2 and anti-VDAC 3 and by matrix assisted laser desorption/ionization time of flight mass spectrometry was used to study the expression of VDAC isoforms in rat, bovine and avian brain mitochondria. We were studying the possibility that differences in the relative expression of VDAC isoforms may be a factor in determining the species-dependent ratio of type A: type B hexokinase binding sites on brain mitochondria. The spots were characterized, and the signal intensities among spots were compared. VDAC1 and VDAC2 were divided into multiple spots. VDAC1 was divided in two spots in two dimensional gels of rat and bovine brains and three spots in avian brains. VDAC2 was separated into three, five and two spots in rat, bovine and avian brains, respectively. The results report charge heterogeneity of VDACs 1 and 2 in the analyzed brains. VDAC1 was the most abundantly expressed of the three isoforms. Moreover the expression of VDAC1 plus VDAC2 was much higher in avian and bovine brains than in rat brains. Avian brain mitochondria showed the highest expression of VDAC1 and the lowest of VDAC2. Bovine brain mitochondria had the highest levels of VDAC2. No VDAC 3 was detected in studied species brains.

ASSUNTO(S)

apoptosis hexoquinase vdac apoptose vdac glicólise hexokinase glycolysis isoformas isoforms

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