The isolation and properties of multiple forms of folate binding protein in cultured KB cells

Archives of Biochemistry and Biophysics
C A LuhrsS P Rothenberg

Abstract

The folate binding proteins (FBPs) of KB cells which were cultured in normal (N) and folate-deficient (D) medium have been characterized. The 200,000 g supernate of lysed cells contained two FBPs which could be separated by DEAE-Bio-Gel A chromatography, indicating that they differ in ionic charge although they could not be separated by gel filtration through Sephadex G-100 (apparent Mr approximately 40,000). Two species of FBP, a major form of apparent Mr approximately 160,000 and a minor form of apparent Mr approximately 40,000, were identified by gel filtration through Sephadex G-150 in the membrane component of the cells after solubilization with Triton X-100. An additional FBP was isolated and purified by affinity chromatography from the medium in which these cells were cultured. By gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the apparent Mr of this FBP was approximately 44,000. The association constants for pteroylglutamic acid of the FBPs in the 200,000 g cell lysate supernate, culture medium, and Triton-solubilized membrane were similar and the relative affinity of folate analogs for the FBP, vis-à-vis pteroylglutamic acid, was similar for all species. An antiserum raised to the purifie...Continue Reading

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Citations

Apr 22, 1988·Biochimica Et Biophysica Acta·M da Costa, S P Rothenberg
Jun 22, 1993·Biochemical Pharmacology·C T Hsueh, B J Dolnick
Jan 1, 1989·Advances in Enzyme Regulation·C B KnightB A Chabner
Jan 4, 1996·Biochimica Et Biophysica Acta·M da Costa, S P Rothenberg
Jan 1, 1996·The Journal of Cell Biology·S RijnbouttG J Strous
Jun 1, 1995·British Journal of Haematology·K ChenD W Essex
Feb 19, 2000·Journal of Drug Targeting·C P LeamonR W Hendren
Jan 30, 2004·The Journal of Nutrition·Michele M Doucette, Victoria L Stevens
Nov 28, 2001·Molecular Pharmacology·M S JhaveriJ B Trepel
Dec 24, 2011·Journal of Proteome Research·Nidhi JaiswalDragan Isailovic

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