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A study of hypoxia sensitive and specific ~(99)Tc~m-MIBI binding protein of mitochondrial membrane in rat myocardium

Objective To investigate the 99 Tc m MIBI binding mechanism independent of the mitochondrial membrane potential Methods The cardiac mitochondrial fragments and heart slices were prepared The techniques of the thin layer chromatography (TLC) and polyacrylamide gel electrophoresis (PAGE) were employed to characterize the specific binding sites of 99 Tc m MIBI Results Two separate peaks of 99 Tc m MIBI binding activity in mitochondrial fragments were identified in TLC One of these binding activity peaks was characterized with a K d of (5 8±1 7) nmol/L and B max of (11 5±3 1) pmol/mg of protein The bound 99 Tc m MIBI could be displaced permanently with 20 μmol/L 99 Tc MIBI and transiently with 50 μmol/L cyclosproin A Three discrete 99 Tc m MIBI labeled proteins of MW about 4 3×10 4, 2 3×10 4 and 1 0×10 4 were revealed in PAGE The clearance of 99 Tc m MIBI could be also induced by hypoxia Conclusions There is a specific 99 Tc m MIBI binding protein which is sensitive to hypoxia metabolism in myocardial mitochondrial membrane with at least two protein subunits of different functions and affinities