Abstract
The lysolecithin extraction procedure originally described by Sadleret al. (1974) has been modified to yield a H+-ATPase with high levels of Pi-ATP exchange activity (400–600 nmol × min−1 × mg−1). This activity is further enhanced (1400–1600 nmol × min−1 × mg−1) following sucrose density gradient centrifugation in the presence of asolectin. This enhancement results in part from a lipid-dependent activation and in part from removal of inactive complexes. The H+ translocating activity of the complex has been determined spectrophotometrically using binding of oxonol VI as an indicator of membrane potential. Pi-ATP exchange, ATP hydrolysis, and oxonol binding are sensitive to energy-transfer inhibitors (oligomycin, rutamycin) and/or uncouplers (DNP, FCCP).
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Hughes, J., Joshi, S., Torok, K. et al. Isolation of a highly active H+-ATPase from beef heart mitochondria. J Bioenerg Biomembr 14, 287–295 (1982). https://doi.org/10.1007/BF00743058
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DOI: https://doi.org/10.1007/BF00743058