Calcium stimulates ATP-Mg/Pi carrier activity in rat liver mitochondria.
Adenine nucleotide transport over the carboxyatractyloside-insensitive ATP-Mg/Pi carrier was assayed in isolated rat liver mitochondria with the aim of investigating a possible regulatory role for Ca2+ on carrier activity. Net changes in the matrix adenine nucleotide content (ATP + ADP + AMP) occur when ATP-Mg exchanges for Pi over this carrier. The rates of net accumulation and net loss of adenine nucleotides were inhibited when free Ca2+ was chelated with EGTA and stimulated when buffered [Ca2+]free was increased from 1.0 to 4.0 microM. The unidirectional components of net change were similarly dependent on Ca2+; ATP influx and efflux were inhibited by EGTA in a concentration-dependent manner and stimulated by buffered free Ca2+ in the range 0.6-2.0 microM. For ATP influx, increasing the medium [Ca2+]free from 1.0 to 2.0 microM lowered the apparent Km for ATP from 4.44 to 2.44 mM with no effect on the apparent Vmax (3.55 and 3.76 nmol/min/mg with 1.0 and 2.0 microM [Ca2+]free, respectively). Stimulation of influx and efflux by [Ca2+]free was unaffected by either ruthenium red or the Ca2+ ionophore A23187. Calmodulin antagonists inhibited transport activity. In isolated hepatocytes, glucagon or vasopressin promoted an increased mitochondrial adenine nucleotide content. The effect of both hormones was blocked by EGTA, and for vasopressin, the effect was blocked also by neomycin. The results suggest that the increase in mitochondrial adenine nucleotide content that follows hormonal stimulation of hepatocytes is mediated by an increase in cytosolic [Ca2+]free that activates the ATP-Mg/Pi carrier.
Nosek
M T
MT
Department of Biology, Tufts University, Medford, Massachusetts 02155.
Dransfield
D T
DT
Aprille
J R
JR
D000227:Adenine Nucleotides / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002122:Calcium Chloride / Q000494:pharmacology*; D002147:Calmodulin / Q000037:antagonists & inhibitors; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D002746:Chlorpromazine / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D005934:Glucagon / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism*; D009355:Neomycin / Q000494:pharmacology; D010710:Phosphates / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013449:Sulfonamides / Q000494:pharmacology; D014268:Trifluoperazine / Q000494:pharmacology; D014667:Vasopressins / Q000494:pharmacology
D000227:Adenine Nucleotides; D002147:Calmodulin; D002352:Carrier Proteins; D007093:Imidazoles; D010710:Phosphates; D013449:Sulfonamides; D014667:Vasopressins; D014268:Trifluoperazine; C031938:calmidazolium; D004533:Egtazic Acid; C017967:W 7; D000255:Adenosine Triphosphate; D005934:Glucagon; D009355:Neomycin; D008274:Magnesium; D002122:Calcium Chloride; D002746:Chlorpromazine