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Related Experiment Videos

Renal brush border membrane bound intrinsic factor.

K S Ramanujam1, S Seetharam, M Ramasamy

  • 1Division of Gastroenterology, Medical College of Wisconsin, Milwaukee, 53226.

Biochimica Et Biophysica Acta
|November 30, 1990
PubMed
Summary

The kidney contains a receptor for intrinsic factor-cobalamin (IF-Cbl), which may prevent vitamin B12 loss in urine. This receptor binds IF-Cbl and contains intrinsic factor, suggesting a role in cobalamin homeostasis.

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Area of Science:

  • Biochemistry
  • Physiology
  • Molecular Biology

Background:

  • Mammalian kidneys possess a receptor for the intrinsic factor (IF)-cobalamin (Cbl) complex.
  • The precise physiological function of this renal receptor in cobalamin homeostasis remains unclear.

Purpose of the Study:

  • To investigate the role of the renal intrinsic factor-cobalamin receptor in cobalamin homeostasis.
  • To identify the presence and characteristics of intrinsic factor in renal membranes.

Main Methods:

  • Binding assays using radiolabeled IF-[57Co]Cbl with renal apical and basolateral membranes.
  • pH 5/EDTA buffer extraction of renal apical membranes.
  • Immunoblotting with anti-IF antiserum to detect intrinsic factor.
  • Analysis of [57Co]Cbl binding and immunoprecipitation in rat urine.

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Main Results:

  • Renal apical membranes exhibit high-affinity binding for IF-[57Co]Cbl and contain endogenous IF/IF-Cbl.
  • A 62 kDa protein, identified as intrinsic factor, was present in renal apical membranes and diminished after pH 5/EDTA wash.
  • The pH 5/EDTA extract containing IF also bound to ileal brush border membranes, promoting ileal Cbl transport.
  • Rat urine showed unsaturated Cbl binding, with only a fraction identified as IF-bound.

Conclusions:

  • The renal IF-Cbl receptor, along with endogenous IF, likely plays a crucial role in retaining cobalamin within the kidney.
  • This mechanism may prevent the urinary excretion of cobalamin, contributing to overall Cbl homeostasis.