Evidence of endoplasmic reticulum-related Ca2+ ATPase in human microvascular endothelial cells.
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.
Bikfalvi
A
A
Inserm U 150 and CNRS URA 184, Hôpital Lariboisière, Paris, France.
Enouf
J
J
Bredoux
R
R
Lompre
A
A
Dupuy
E
E
Bourdeau
N
N
Levy-Toledano
S
S
Tobelem
G
G
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology; D015152:Blotting, Northern; D015153:Blotting, Western; D000252:Calcium-Transporting ATPases / Q000235:genetics / Q000378:metabolism*; D015342:DNA Probes; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D004730:Endothelium, Vascular / Q000201:enzymology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D011233:Precipitin Tests; D011817:Rabbits
D015342:DNA Probes; D000252:Calcium-Transporting ATPases