PMID: 6540119Jul 25, 1984Paper

Chromatofocussing and centrifugal reconstitution as tools for the separation and characterization of the Na+-cotransport systems of the brush-border membrane

Biochimica Et Biophysica Acta
J T LinA Stroh

Abstract

Chromatofocussing was used for the separation of brush-border membrane proteins from calf kidney into 4 or 5 fractions over the pH range 4.0 to 7.4. These fractions were reconstituted into proteoliposomes by gradient centrifugation. Determination of the sodium-dependent solute uptake by proteoliposomes reconstituted from different chromatofocussed fractions showed that the sodium-D-glucose cotransport system was present in the fraction eluted between pH 5.3 and 5.8, and that the sodium-phosphate cotransport was present in fractions eluted between pH 4.6 and 5.3 and between pH 5.8 and 6.6, sodium-alanine cotransport could be detected in almost all fractions. Marker enzymes of the brush-border membrane, such as alkaline phosphatase, gamma-glutamyltransferase and aminopeptidase M etc. were also found to be eluted at pH 7.0-7.4, 4.0-4.1 and 5.6-5.8, respectively. These results suggest that chromatofocussing is a promising tool for the separation of membrane proteins and for pre-purification of the sodium-D-glucose cotransport system. It can be further concluded that the sodium-dependent phosphate transport across the brush-border membrane is not dependent upon alkaline phosphatase activity.

References

Oct 19, 1979·Biochimica Et Biophysica Acta·J T LinR Kinne
Mar 25, 1975·Biochimica Et Biophysica Acta·A Helenius, K Simons
Jan 1, 1982·Physiological Reviews·A J Kenny, S Maroux
Aug 1, 1981·Archives of Biochemistry and Biophysics·D BrèthesJ Chevallier
Jul 15, 1980·The Journal of Membrane Biology·H Murer, R Kinne
Jun 14, 1982·Biochimica Et Biophysica Acta·G ToggenburgerG Semenza
Aug 16, 2011·Cancer Research·Edgar S Díaz-CruzPriscilla A Furth

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Citations

Nov 1, 1989·The Journal of Membrane Biology·J P Wehrle, P L Pedersen
Nov 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·B E Peerce, E M Wright
Jan 1, 1985·Annals of the New York Academy of Sciences·J T Lin
Aug 1, 1987·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·D M Scott
Dec 1, 1993·British Journal of Pharmacology·J F KlehaA Johns
Aug 1, 1986·Kidney International·H Murer, P Gmaj

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