Evidence of endoplasmic reticulum-related Ca2+ ATPase in human microvascular endothelial cells |
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Authors: | A Bikfalvi J Enouf R Bredoux A Lompre E Dupuy N Bourdeau S Levy-Toledano G Tobelem |
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Institution: | Inserm U 150 and CNRS URA 184, H?pital Lariboisière, Paris, France. |
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Abstract: | We have demonstrated by immunological and molecular methods the presence of a reticulum endoplasmic-related Ca2+-ATPase in human omental microvascular endothelial cells (HOME cells). HOME cells reacted positively with a previously characterized sarcoplasmic reticulum Ca2+-ATPase antibody as demonstrated by indirect immunofluorescence. Western blotting revealed that the antibody recognized a 95-100 kDa protein. 35S-Metabolic labeling led to the detection of a similar protein with which the purified sarcoplasmic reticulum Ca2+-ATPase competed. Dot-blotting experiments indicated that a substantial amount of Ca2+-ATPase was present in HOME cell membranes. In addition, Northern blot analysis using a cDNA probe from cardiac sarcoplasmic reticulum showed the presence of mRNA species of 4-kb. As these experiments were conducted in comparison with cell types with well-defined Ca2+-ATPase in HOME cells. |
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