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. 1988 Jan;85(1):46–50. doi: 10.1073/pnas.85.1.46

Isolation and structural characterization of a cDNA clone encoding rat gastric intrinsic factor.

B K Dieckgraefe 1, B Seetharam 1, L Banaszak 1, J F Leykam 1, D H Alpers 1
PMCID: PMC279478  PMID: 3422425

Abstract

Rat intrinsic factor (IF) has been purified and proteolytic fragments were sequenced. A cDNA library was constructed from size-enriched gastric poly(A)+ RNA and screened for IF-positive clones by antibody and synthetic oligodeoxynucleotide probe hybridization. An IF clone was isolated and sequenced, revealing a predicted primary amino acid sequence in the coding region of 421 amino acids and a putative signal sequence of 22 amino acids. The primary translation product of IF produced in a cell-free translation system displayed cobalamin (Cbl)-binding activity without proteolytic processing or glycosylation. The amino-terminal region of IF showed significant secondary structural and hydropathic homologies with the nucleotide-binding domain in NAD-dependent oxidoreductases. Alignment of the first 80 residues of IF, following the signal peptide, demonstrated homology with the nucleotide-binding domain of cytoplasmic malate dehydrogenase. Based on these data, we propose a model of IF tertiary structure in which the Cbl-binding domain resides in the NH2-terminal half of the protein.

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Selected References

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