Interaction of lipids with intestinal brush border membrane preparations

1982 ◽  
Vol 60 (9) ◽  
pp. 904-909 ◽  
Author(s):  
P. Proulx ◽  
J. McNeil ◽  
I. Brglez ◽  
D. G. Williamson
Keyword(s):  
Brush Border ◽  
Protein Concentration ◽  
Brush Border Membrane ◽  
Divalent Cations ◽  
Monovalent Cations ◽  
Intestinal Brush Border Membrane ◽  
Intestinal Brush Border ◽  
Ph Range ◽  
Assay Conditions ◽  
Lipid Structures

Conditions for uptake of lipids by rabbit intestinal brush border membrane preparations were investigated. A variety of lipids were found to be incorporated, including choline and ethanolamine phosphatides as well as cholesterol, diglyceride, and fatty acid. The incorporation of those lipids tested was enhanced by Ca2+ and other divalent cations but not by monovalent cations. The optimal Ca2+ concentration was approximately 10 mM. The uptake varied with lipid and membrane protein concentration and proceeded at rates which were too rapid to measure under several assay conditions tried. Incorporations were decreased substantially outside the pH range of 6.5–8.0. The effect of one lipid, phosphatidylcholine, on the structural appearance of the membrane fraction was examined by electron microscopy. No free or surface-bound lipid structures could be detected and the membrane fractions appeared to be unchanged after uptake.

Divalent cations and vitamin B12 uptake by intestinal brush-border membrane vesicles

1980 ◽  
Vol 239 (6) ◽  
pp. G452-G456
Author(s):  
R. C. Beesley ◽  
C. D. Bacheller
Keyword(s):  
Vitamin B12 ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Divalent Cations ◽  
Intrinsic Factor ◽  
Membrane Vesicles ◽  
Brush Border Membrane Vesicles ◽  
Tetraacetic Acid ◽  
Intestinal Brush Border Membrane ◽  
Intestinal Brush Border

Brush-border membrane vesicles from hamster intestine were employed to investigate uptake (binding) of vitamin B12 (B12). Ileal vesicles took up 25 times more B12 than did jejunal vesicles. Uptake of B12 by ileal vesicles was dependent on intrinsic factor (IF) and required Ca2+. Increasing the Ca2+ concentration caused an increase in uptake of B12 reaching a maximum at approximately 8 mM Ca2+. At high Ca2+ concentrations, 6–8 mM, Mg2+ had little effect on uptake of B12. At low Ca2+ concentrations, up to 2 mM, Mg2+ stimulated B12 uptake. Mg2+, Mn2+, and, to a lesser extent, Sr2+ stimulated Ca2+-dependent B12 uptake, but Zn2+, Ba2+, Na+, K+, and La3+ did not. B12 was apparently not metabolized and was bound as IF-B12 complex, which could be removed with (ethylenedinitrilo)tetraacetic acid (EDTA). Our results suggest that two types of divalent cation reactive sites are involved in binding of IF-B12. One is Ca2+ specific. The other is less specific reacting with Mg2+, Mn2+, Sr2+, and perhaps Ca2+ itself, thereby stimulating Ca2+-dependent binding of IF-B12 to its ileal receptor.

Reconstitution of intestinal Na(+)-phosphate cotransporter

1993 ◽  
Vol 264 (4) ◽  
pp. G609-G616 ◽  
Author(s):  
B. E. Peerce ◽  
M. Cedilote ◽  
S. Seifert ◽  
R. Levine ◽  
C. Kiesling ◽  
...  
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Phosphate Uptake ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Preparative Scale ◽  
Intestinal Brush Border Membrane ◽  
Intestinal Brush Border ◽  
Ph Range

The rabbit intestinal brush-border membrane Na(+)-phosphate cotransporter was purified from sodium dodecyl sulfate (SDS)-brush-border membrane vesicles (BBMV) protein (SDS-treated Ca(2+)-precipitated BBMV) by a three-column chromatography protocol. The purification included a preparative scale chromatofocusing chromatography column over the pH range from 7.4 to 4 after solubilization in 3-[(3-cholamidopropyl)-diamethylammonia]-1-propanesulfonate (CHAPS), a chromatofocusing column over the pH range from 5.6 to 4 after solubilization in n-octyl glucoside, and gel filtration chromatography on a Sephacryl S-200 column. Verification of Na(+)-phosphate cotransporter purification involved substrate affinities, substrate stoichiometry, and inhibitor sensitivity after proteoliposome reconstitution and SDS-polyacrylamide gel electrophoresis (PAGE). After gel filtration Na(+)-dependent phosphate uptake was 3,300-fold enriched compared with the cell homogenate. A single 130-kDa polypeptide was visualized by SDS-PAGE under reducing conditions using silver stain. The coenrichment of this 130-kDa polypeptide and proteoliposome reconstituted Na(+)-dependent phosphate uptake suggest that the intestinal brush-border membrane Na(+)-phosphate cotransporter has been purified and proteoliposome reconstituted.

Zinc binding in intestinal brush-border membrane isolated from pig

1991 ◽  
Vol 1063 (1) ◽  
pp. 51-59 ◽  
Author(s):  
Frédérique Tacnet ◽  
Don W. Watkins ◽  
Pierre Ripoche
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Zinc Binding ◽  
Intestinal Brush Border Membrane ◽  
Intestinal Brush Border

scholarly journals Class B type I scavenger receptor is responsible for the high affinity cholesterol binding activity of intestinal brush border membrane vesicles

2007 ◽  
Vol 1771 (9) ◽  
pp. 1132-1139 ◽  
Author(s):  
Eric D. Labonté ◽  
Philip N. Howles ◽  
Norman A. Granholm ◽  
Juan C. Rojas ◽  
Joanna P. Davies ◽  
...  
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Scavenger Receptor ◽  
Membrane Vesicles ◽  
Binding Activity ◽  
Type I ◽  
Intestinal Brush Border Membrane ◽  
High Affinity ◽  
Intestinal Brush Border ◽  
Class B
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