title: ERp60 does not substitute for protein disulphide isomerase as the beta-subunit of prolyl 4-hydroxylase.
abstract: Prolyl 4-hydroxylase (EC 1.14.11.2) catalyses the formation of 4-hydroxyproline in collagens. The vertebrate enzymes are alpha 2 beta 2 tetramers while the Caenorhabditis elegans enzyme is an alpha beta dimer. The beta-subunit is identical to protein disulphide isomerase (PDI), a multifunctional endoplasmic reticulum luminal polypeptide. ERp60 is a PDI isoform that was initially misidentified as a phosphatidylinositol-specific phospholipase C. We report here on the cloning and expression of the human and Drosophila ERp60 polypeptides. The overall amino acid sequence identity and similarity between the processed human ERp60 and PDI polypeptides are 29% and 56% respectively, and those between the Drosophila ERp60 and human PDI polypeptides 29% and 55%. The two ERp60 polypeptides were found to be similar to human PDI within almost all their domains, the only exception being the extreme C-terminal region. Nevertheless, when the human or Drosophila ERp60 was expressed in insect cells together with an alpha-subunit of human prolyl 4-hydroxylase, no tetramer was formed and no prolyl 4-hydroxylase activity was generated in the cells. Additional experiments with hybrid polypeptides in which the C-terminal regions had been exchanged between the human ERp60 and PDI polypeptides demonstrated that the differences in the C-terminal region are not the only reason for the lack of prolyl 4-hydroxylase tetramer formation by ERp60.
authors.affiliation: Collagen Research Unit, Biocenter, University of Oulu, Finland.
authors.forename: P
authors.initials: P
authors.lastname: Koivunen
authors.forename: T
authors.initials: T
authors.lastname: Helaakoski
authors.forename: P
authors.initials: P
authors.lastname: Annunen
authors.forename: J
authors.initials: J
authors.lastname: Veijola
authors.forename: S
authors.initials: S
authors.lastname: Räisänen
authors.forename: T
authors.initials: T
authors.lastname: Pihlajaniemi
authors.forename: K I
authors.initials: KI
authors.lastname: Kivirikko
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D037282:Calreticulin; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D004330:Drosophila / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D019143:Evolution, Molecular; D006801:Humans; D007535:Isomerases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011393:Procollagen-Proline Dioxygenase / Q000737:chemistry* / Q000378:metabolism; D019704:Protein Disulfide-Isomerases; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D002135:Calcium-Binding Proteins; D037282:Calreticulin; D018076:DNA, Complementary; D010455:Peptides; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D011393:Procollagen-Proline Dioxygenase; D007535:Isomerases; D019704:Protein Disulfide-Isomerases