The primary structure of human hemopexin deduced from cDNA sequence: evidence for internal, repeating homology

• Published in: Nucleic acids research 1985-06, Vol.13 (11), p.3841-3859
• Main Authors: Altruda, Fiorella, Poli, Valeria, Restagno, Gabriella, Argos, Patrick, Cortese, Riccardo, Silengo, Lorenzo
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Applied sciences; Base Sequence; cDNA; Cloning, Molecular; DNA - analysis; DNA Restriction Enzymes; Exact sciences and technology; hemopexin; Hemopexin - genetics; Humans; Nucleic Acid Hybridization; nucleotide sequence; Other techniques and industries; Protein Conformation; Repetitive Sequences, Nucleic Acid
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/13.11.3841

We have cloned and analyzed a cDNA containing the coding sequence for human hemopexin. We have first identified, by immunological screening of 30.000 colonies of a liver cDNA library in the expression vector pEX1, a clone carrying an insert 1170 base pairs long that shows 100% homology with a known human hemopexin peptide. The complete sequence coding for hemopexin was isolated from a liver cDNA library in the vector pAT218. The DNA insert of 1523 base pairs shows an open reading frame coding for 439 amino acids, a 3′ noncoding region of 159 nucleotides long, followed by a poly(A) tail. The insert spans the entire coding region and from which the primary structure of the protein was deduced. By computer assisted analysis of the amino acid sequence, it was possible to recognize a core unit, of about 45 amino acids, which is repeated 8 or possibly even 10 fold along the polypeptide chain. This feature suggests that the gene might have evolved through a series of duplications. This characteristic, together with prediction of secondary structure, suggest a rough model for the tridimensional folding that allows some speculations on the function of hemopexin. Blot hybridization of total RNA from human liver with nick translated hemopexin cDNA detected a message of about 1600 nucleotides. Southern blot experiments to identify the hemopexin gene (s) suggest that it is not a large multi-gene family, but that there is only one or at most a few genes in the human genome.