Properties of a folate binding protein (FBP) isolated from porcine kidney.
A specific high-affinity folate binding protein (FBP) that binds folic acid and folic acid derivatives and that was previously identified in porcine kidney has been purified 50,000-fold using the technique of affinity chromatography. The FBP had a molecular weight of 38,500 daltons and did not appear to aggregate in solution, as has been reported to be the case with folate binding protein from milk. At pH 7.6, the Ka was at least 5 X 10(12)M-1. At pH values greater than 9.5 or less than 5, the binding dramatically decreased. The specificity was determined by an isotopic dilution technique using [3H]folic acid and folic acid analogs and derivatives. The FBP reacted more rapidly with unsubstituted folates, and the number of glutamic acid moieties (N greater than or equal to 1) did not influence binding. Binding of folic acid to the FBP was unaffected by a variety of anions and cations, and 8 M urea, but was disrupted by 6 M guanidine hydrochloride. Proteolytic enzymes irreversibly destroyed binding affinity, but RNase, DNase, phospholipase and neuraminidase had no effect.
D000818:Animals; D001667:Binding, Competitive; D001798:Blood Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D005260:Female; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000032:analysis*; D008099:Liver / Q000378:metabolism; D008894:Milk Proteins / Q000378:metabolism; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface; D013379:Substrate Specificity; D013552:Swine / Q000378:metabolism
D001798:Blood Proteins; D002352:Carrier Proteins; D058973:Folate Receptors, GPI-Anchored; D008894:Milk Proteins; D011257:Pregnancy Proteins; D011956:Receptors, Cell Surface; D005492:Folic Acid