Characterization of Na(+)-Ca2+ exchange activity in plasma membrane vesicles from postmortem human brain.
Procedures were developed for measurement of Na+/Ca2+ exchange in resealed plasma membrane vesicles from postmortem human brain. The vesicle preparation method permits use of stored frozen tissue with minimal processing required prior to freezing. Vesicles prepared in this manner transport Ca2+ in the presence of a Na+ gradient. The kinetic characteristics of the Na+/Ca2+ exchange process were determined in membrane vesicles isolated from hippocampus and cortex. The Kact for Ca2+ was estimated to be 32 microM for hippocampal and 17 microM for cortical tissue. The maximal rate of Ca2+ uptake (Vmax) was 3.5 nmol/mg protein/15 sec and 3.3 nmol/mg protein/15 sec for hippocampal and cortical tissue, respectively. Exchange activity was dependent on the Na+ gradient, and was optimal in the high pH range. Therefore, membranes in which Na(+)-dependent Ca2+ transport activity is preserved can be isolated from postmortem human brain and could be used to determine the influence of pathological conditions on this transport system.
Hoel
G
G
Department of Pharmacology and Toxicology, University of Kansas, Lawrence 66047.
Michaelis
M L
ML
Freed
W J
WJ
Kleinman
J E
JE
D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D006096:Gramicidin / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D011180:Postmortem Changes; D012964:Sodium / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013570:Synaptic Membranes / Q000187:drug effects / Q000378:metabolism*; D014021:Tissue Preservation / Q000379:methods
D002352:Carrier Proteins; D019831:Sodium-Calcium Exchanger; D006096:Gramicidin; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase; D002118:Calcium