Isolation and characterization of two species of receptor which bind intrinsic factor.
The intrinsic factor receptor from guinea pig ilea has been characterized following purification by affinity chromatography. The purified receptor complexed to intrinsic factor-cobalamin (holo-receptor) had an anhydrous molecular weight of 680,000, a Stokes radius of 10.9 nm, and a sedimentation coefficient of 15.1 S. In contrast, unsaturated receptor (apo-receptor) resolved into distinct "large" and "small" molecular species having, respectively, (i) molecular weights of 700,000 and 350,000; (ii) Stokes radii of 11.1 and 7.06 nm; (iii) sedimentation coefficients of 15.5 S and 11.9 S; (iv) association constants for binding the intrinsic factor-cobalamin complex of 7.3 and 2.5 X 10(10) liters mol-1; and (v) two isoproteins for the larger species (pI's 4.05 and 4.80), and one isoprotein for the smaller species (pI 4.90). A rabbit anti-receptor serum gave only one precipitation line in immunodiffusion against either the large or the small receptor, and each one of these two lines fused completely with the one of two lines which formed with the purified preparation containing both receptor species. Autoradiography of the precipitin lines obtained when the receptor was coupled to intrinsic factor-cyano[57Co]cobalamin demonstrated that both species of receptor were functional. The reaction of complete immunologic identity, the similar electrical properties and similar kinetics for binding to intrinsic factor, and the observed molecular weight differences indicate that the small and large apo-receptors are chemically interrelated, and suggest that the large receptor may consist of two small functional proteins.
D000818:Animals; D001059:Apoproteins / Q000302:isolation & purification; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D006168:Guinea Pigs; D007082:Ileum / Q000032:analysis; D007120:Immunochemistry; D007413:Intestinal Mucosa / Q000032:analysis*; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D011487:Protein Conformation; D011956:Receptors, Cell Surface / Q000302:isolation & purification*; D018000:Receptors, Peptide
D001059:Apoproteins; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; C028199:intrinsic factor receptor