Molecular cloning of the beta-subunit of human prolyl 4-hydroxylase. This subunit and protein disulphide isomerase are products of the same gene.
Prolyl 4-hydroxylase (EC 1.14.11.2), an alpha 2 beta 2 tetramer, catalyses the formation of 4-hydroxyproline in collagens by the hydroxylation of proline residues in peptide linkages. We report here the isolation of cDNA clones coding for the beta-subunit of prolyl 4-hydroxylase from a human hepatoma lambda gt11 library and a corresponding human placenta library. Five overlapping clones covering all the coding sequences and almost all the non-coding sequences were characterized. The size of the mRNA hybridizing with these clones in Northern blotting is approximately 2.5 kb. The clones encode a polypeptide of 508 amino acid residues, including a signal peptide of 17 amino acids. These human sequences were found to be very similar to those recently reported for rat protein disulphide isomerase (EC 5.3.4.1). The degree of homology between these two proteins was 84% at the level of nucleotide sequences or 94% at the level of amino acid sequences. Southern blot analyses of human genomic DNA with a cDNA probe for the beta-subunit indicated the presence of only one gene containing these sequences. The product of a single gene thus appears to possess two different enzymatic functions depending on whether it is present in cells in monomer form or in the prolyl 4-hydroxylase tetramer.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D004262:DNA Restriction Enzymes; D005260:Female; D005796:Genes; D006801:Humans; D007535:Isomerases / Q000235:genetics*; D008113:Liver Neoplasms; D008114:Liver Neoplasms, Experimental; D046911:Macromolecular Substances; D009693:Nucleic Acid Hybridization; D010920:Placenta / Q000201:enzymology; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics*; D019704:Protein Disulfide-Isomerases
D046911:Macromolecular Substances; D004247:DNA; D011393:Procollagen-Proline Dioxygenase; D004262:DNA Restriction Enzymes; D007535:Isomerases; D019704:Protein Disulfide-Isomerases