Literature DB >> 12576057

In situ assay of the intramitochondrial enzymes: use of alamethicin for permeabilization of mitochondria.

Irina S Gostimskaya1, Vera G Grivennikova, Tatyana V Zharova, Lora E Bakeeva, Andrei D Vinogradov.   

Abstract

The channel-forming antibiotic alamethicin was used to permeabilize mitochondrial membranes for the low molecular mass hydrophilic substrates NADH and ATP. Alamethicin-treated mitochondria show high rotenone-sensitive NADH oxidase, NADH-quinone reductase, and oligomycin-sensitive and carboxyatractylate-insensitive ATPase activities. Alamethicin does not affect Complex I and ATPase activities in inside-out submitochondrial particles. Permeabilized mitochondria quantitatively retain their aconitase and iso-citrate dehydrogenase activities. Electron microscopy of alamethicin-treated mitochondria reveals no disruption of their outer and inner membranes. From the results obtained it is recommended, that alamethicin be used for the in situ catalytic assay of intramitochondrially located enzymes. Copyright 2003 Elsevier Science (USA)

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Year:  2003        PMID: 12576057     DOI: 10.1016/s0003-2697(02)00534-1

Source DB:  PubMed          Journal:  Anal Biochem        ISSN: 0003-2697            Impact factor:   3.365


  37 in total

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