Purification of brush border membrane vesicles from rat renal cortex by size-exclusion chromatography.
Size-exclusion chromatography with controlled pore glass (CPG) was used in the further purification of renal brush border membrane vesicles (BBMV) isolated by the Ca precipitation method. The BBMV obtained had an almost spherical shape and their average diameter was about 95 nm in isotonic solution. The specific activities of alkaline phosphate and leucine aminopeptidase in the BBMV preparation were increased 18- and 17-fold, respectively, over those in the crude homogenate. The uptake of D-glucose by the purified BBMV in the presence of a sodium gradient reached 8.53 nmol/mg protein at 20 s. These results indicate that CPG chromatography is suitable procedure by which to obtain purified renal BBMV of homogenous size and with high specific marker enzyme activity for use in the study of membrane transport.
Nagasawa
M
M
Department of Medicine, Juntendo University School of Medicine, Tokyo, Japan.
Koide
H
H
Ohsawa
K
K
Hoshi
T
T
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002845:Chromatography / Q000379:methods*; D005947:Glucose / Q000378:metabolism; D007672:Kidney Cortex / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D008871:Microvilli / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure*; D051381:Rats; D011919:Rats, Inbred Strains
D005947:Glucose