Purification and Characterization of PMC16 Nanoparticles from Rat Heart Mitochondria

Up to recently, a not-too-long list of the mitochondrial porphyrin-binding proteins (PBP) was strictly limited to well known heme-containing molecules like cytochromes or peroxidases [1]. In external membrane of mammalian mitochondria, however, some minor group of proteins with a marked capability for selective binding of porphyrin K and its derivatives but with no direct link to either heme or any other porphyrin in situ has been found lately [2]. Both functional role and structural peculiarities of these specific membrane proteins remains obscure which requires a reliable method to isolate / purify these unique compounds for their further study.The porphyrin – engaging ligands look suitable for affinity chromatography of PBP and related compounds. It is not easy, nonetheless, to reach a perfect protein – ligand coupling as long as the whole stationary phase molecular architecture does not allow to make it stoichiometrically (nanotopologically) correct. Due to this, a number of technical problems are found to be the hard obstacles on a way to the PBP affinity chromatography developments [3].To solve these problems, we have proposed an affinity chromatography technique based on application of a new stationary phase possessing the porphyrin domain as a ligand attached to the complex epoxy[cyclohecyl]C60-fullerene spacer immobilized firmly on an agarose gel matrix. In the best of our knowledge, this is the first report ever on the fullerene nanostructure use to optimize a protein-porphyrin recognition in affinity coupling.