[Preparation of inside-out vesicles from red blood cell membranes and measurement for its Ca2+ uptake].
Sealed, inside-out red cell membrane vesicles (IOV) from rats were prepared by a modified method of Steck (1974). The preparation contained 70% of IOV. In the presence of ATP and Mg2+, these vesicles actively took up Ca2+ with a high efficiency and reproducibility. The intraassay coefficient of variance (C.V.) was 3.6% and the interassay C.V. was 13.58%. The active Ca2+ transport reached the maximal level in 10 minutes of incubation, and linearly correlated with the IOV concentration. The Ca2+ uptake rate in Wistar rats was 9.39 +/- 1.28 nmol/mg IOV protein/min. IOV rapidly lost Ca2+ when A23187 was added. The active transport was stimulated by calmodulin, and inhibited by trifluoperazine.
Luo
H
H
Institute of Basic Medical Sciences, Beijing.
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D002147:Calmodulin / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D014268:Trifluoperazine / Q000494:pharmacology
D015220:Calcium Channels; D002147:Calmodulin; D014268:Trifluoperazine; D000001:Calcimycin; D000255:Adenosine Triphosphate; D002118:Calcium