ArticleMolecular and cellular biochemistry1996
The uptake of calcium by isolated chromaffin granules of the adrenal medulla.
Article in Molecular and cellular biochemistry, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Bovine chromaffin secretory granules were purified by isopycnic Metrizamide gradient centrifugation and their Ca2+ sequestration pathways were characterized. The rate of Ca2+ sequestration at 37 degrees C was first order, with a maximal uptake of 26.9 +/- 0.46 (mean +/- S.D., n = 3) nmol Ca2+/mg protein and a first order rate constant (k) of 0.046 +/- 0.002 min-1. At 4 degrees C the rate of uptake was substantially attenuated, with only 2.47 +/- 0.2 (mean +/- S.D, n = 3) nmol Ca2+/mg protein sequestered in 60 min. Ca2+ sequestration was 93% inhibited by 180 mM NaCl [I50% of 78.7 +/- 9.3 mM NaCl (mean +/- S.D., n = 11)] but only slightly inhibited by KCl or MgCl2. Ca2+ sequestration was not stimulated by incubation with MgATP but was inhibited by 57% after incubation with 30 microM monensin. Ca2+ sequestration was dependent on extravesicular Ca2+ with half-maximal sequestration at pCa2+ 6.81 +/- 0.028 (mean +/- S.D., n = 3). Sequestered Ca2+ could be exchanged with external 45Ca2+, the exchange rate was first order (k of 0.042 +/- 0.004: mean +/- S.D., n = 3) and saturated at 27.7 +/- 1.1 nmol Ca2+/mg (mean +/- S.D., n = 3). The Ca2+/Ca2+ exchange system was totally inhibited by NaCl or KCl but only slightly by MgCl2. About 75% of sequestered 45Ca2+ could be released by incubation with NaCl, but only 8% was released by incubation with KCl. Half-maximal release of sequestered 45Ca2+ required 69.3 +/- 12.2 mM NaCl (mean +/- S.D., n = 3). The Na+-induced release of sequestered 45Ca2+ was rapid, t0.5 of 2.80 +/- 0.63 min (mean +/- S.D., n = 3) and inhibited at 4 degrees C. The concurrent incubation of chromaffin granules with 45Ca2+ and either annexin proteins V or VI resulted in attenuated uptake of 45Ca2+. These results suggest that Ca2+ uptake in adrenal chromaffin granules is regulated by Na+ and Ca2+ gradients and also possibly by annexins V and VI.
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