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Cubilin P1297L mutation associated with hereditary megaloblastic anemia 1 causes impaired recognition of intrinsic factor–vitamin B12 by cubilin by Mette Kristiansen, Maria Aminoff, Christian Jacobsen, Albert de la Chapelle, Ralf Krahe, Pierre J. Verroust, and Søren K. Moestrup Blood Volume 96(2):405-409 July 15, 2000 ©2000 by American Society of Hematology Mammalian expression of the IF–Cbl-binding region of wild-type and FM1 mutant cubilin.(A) The modular protein organization of human cubilin (460 kd) and the IF–Cbl-binding region (cubilin CUB domains 5-8, amino acid 928-1386) expressed in CHO cells stably t... Mette Kristiansen et al. Blood 2000;96:405-409 ©2000 by American Society of Hematology Elution profiles of IF–Cbl-purified wild-type and FM1 mutant CUB 5-8 proteins.Nonreducing SDS–PAGE (silver staining) of the eluted fractions. Mette Kristiansen et al. Blood 2000;96:405-409 ©2000 by American Society of Hematology Proteolytic digestion of the wild-type and FM1 mutant CUB 5-8 proteins.Analysis by reducing SDS–PAGE and silver staining. Mette Kristiansen et al. Blood 2000;96:405-409 ©2000 by American Society of Hematology Surface plasmon resonance analysis of the binding of IF–Cbl flowing along BIAcore CM5 sensor chips with immobilized wild-type or FM1 mutant form of CUB 5-8.(A) The IF-Cbl binding to the wild-type CUB 5-8 protein with a flow concentration of 10, 20, and 50 n... Mette Kristiansen et al. Blood 2000;96:405-409 ©2000 by American Society of Hematology Uptake of 125I–IF–Cbl in cubilin-expressing epithelial cells and inhibition with the wild-type and FM1 mutant CUB 5-8 proteins.(A) Time-course for cell association (•) and degradation (○) of iodine 125–IF–Cbl. Mette Kristiansen et al. Blood 2000;96:405-409 ©2000 by American Society of Hematology Effect of wild-type and FM1 mutant CUB 5-8 proteins on the total uptake of 125I–IF–Cbl in epithelial cells.Values are the mean ± 1 SD of triplicate experiments and are shown as the percentage of uptake in relation to the control uptake of iodine 125–IF–Cbl ... Mette Kristiansen et al. Blood 2000;96:405-409 ©2000 by American Society of Hematology