Characteristics of exogenous lipid uptake by renal and intestinal brush-border membrane vesicles.
The transfer of radioactive phosphatidylcholine (PC*) from liposomes to rabbit jejunal and renal brush-border membrane vesicles (BBMVs) was measured with a fast-sampling, rapid-filtration apparatus. PC* uptake by jejunal and renal BBMVs was favoured when liposomes were made from soybean phosphatidylcholine (azolectin, AZO), whereas PC* uptake could not be quantitatively assessed from egg yolk phosphatidylcholine (PC) liposomes even after a 22-h period of incubation. The increased turbidity of BBMV dispersion following the addition of CaCl2 or HCl to AZO-treated BBMVs suggested that negatively charged lipids and phosphatidylethanolamine are transferred during the process. These data and the analysis of PC*-uptake time measurements, using an algorithm simulating aggregation phenomena, indicated that the reaction mechanism involved liposome aggregation to BBMVs rather than specific lipid transfer. The constants of the dimerization reaction between AZO liposomes and BBMVs were evaluated to be 0.016 +/- 0.006 min-1 for jejunal and 0.095 +/- 0.02 min-1 for renal preparations. IntraveSICULAR D-ASPartic acid accumulation in the presence of a NA+ gradient indicated that vesicles were still closed after coincubation with liposomes. In contrast, 70-85% of rabbit jejunal and renal Na(+)-D-glucose cotransporter activities were lost after overnight incubation with either AZO liposomes or buffered solution. Further, H(+)-ATPase activity in rabbit renal BBMVs largely decreased after coincubation with AZO liposomes, while brush-border membrane associated enzymes remained stable. These results demonstrate that coincubation of BBMV with liposomes of different composition may represent a useful approach to study the influence of lipidic environment on various membrane protein functions.
Bellemare
F
F
Department of Physiology, Faculty of Medicine, Université de Montréal, PQ, Canada.
Noël
J
J
Malo
C
C
D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism*; D018699:Coated Vesicles / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism*; D050356:Lipid Metabolism; D008081:Liposomes; D006180:Proton-Translocating ATPases / Q000378:metabolism; D011817:Rabbits
D008081:Liposomes; D006180:Proton-Translocating ATPases