Prolyl 3-hydroxylase 1, enzyme characterization and identification of a novel family of enzymes

Janice Vranka, Lynn Sakai, Hans Peter Bächinger

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

The collagen prolyl hydroxylases are enzymes that are required for proper collagen biosynthesis, folding, and assembly. They reside within the endoplasmic reticulum and belong to the group of 2-oxoglutarate and iron-dependent dioxygenases. Although prolyl 4-hydroxylase has been characterized as an α2β2 tetramer in which protein disulfide isomerase is the β subunit with two different α subunit isoforms, little is known about the enzyme prolyl 3-hydroxylase (P3H). It was initially characterized and shown to have an enzymatic activity distinct from that of prolyl 4-hydroxylase, but no amino acid sequences or genes were ever reported for the mammalian enzyme. Here we report the characterization of a novel prolyl 3-hydroxylase enzyme isolated from embryonic chicks. The primary structure of the enzyme, which we now call P3H1, demonstrates that P3H1 is a member of a family of prolyl 3-hydroxylases, which share the conserved residues present in the active site of prolyl 4-hydroxylase and lysyl hydroxylase. P3H1 is the chick homologue of mammalian leprecan or growth suppressor 1. Two other P3H family members are the genes previously called MLAT4 and GRCR In this study we demonstrate prolyl 3-hydroxylase activity of the purified enzyme P3H1 on a full-length procollagen substrate. We also show it to specifically interact with denatured collagen and to exist in a tight complex with other endoplasmic reticulum-resident proteins. Immunohistochemistry with a monoclonal antibody specific for chick P3H1 localizes P3H1 specifically to tissues that express fibrillar collagens, suggesting that other P3H family members may be responsible for modifying basement membrane collagens.

Original languageEnglish (US)
Pages (from-to)23615-23621
Number of pages7
JournalJournal of Biological Chemistry
Volume279
Issue number22
DOIs
StatePublished - May 28 2004

Fingerprint

Prolyl Hydroxylases
Enzymes
Collagen
Endoplasmic Reticulum
2-Oxoglutarate 5-Dioxygenase Procollagen-Lysine
Genes
Fibrillar Collagens
Protein Disulfide-Isomerases
Dioxygenases
Procollagen
Biosynthesis
Basement Membrane
2-oxoglutarate 3-dioxygenase proline
Amino Acid Sequence
Catalytic Domain
Protein Isoforms
Iron
Immunohistochemistry
Monoclonal Antibodies
Tissue

ASJC Scopus subject areas

  • Biochemistry

Cite this

Prolyl 3-hydroxylase 1, enzyme characterization and identification of a novel family of enzymes. / Vranka, Janice; Sakai, Lynn; Bächinger, Hans Peter.

In: Journal of Biological Chemistry, Vol. 279, No. 22, 28.05.2004, p. 23615-23621.

Research output: Contribution to journalArticle

@article{78715a10cf6649bea5d5b9f11260fe5b,
title = "Prolyl 3-hydroxylase 1, enzyme characterization and identification of a novel family of enzymes",
abstract = "The collagen prolyl hydroxylases are enzymes that are required for proper collagen biosynthesis, folding, and assembly. They reside within the endoplasmic reticulum and belong to the group of 2-oxoglutarate and iron-dependent dioxygenases. Although prolyl 4-hydroxylase has been characterized as an α2β2 tetramer in which protein disulfide isomerase is the β subunit with two different α subunit isoforms, little is known about the enzyme prolyl 3-hydroxylase (P3H). It was initially characterized and shown to have an enzymatic activity distinct from that of prolyl 4-hydroxylase, but no amino acid sequences or genes were ever reported for the mammalian enzyme. Here we report the characterization of a novel prolyl 3-hydroxylase enzyme isolated from embryonic chicks. The primary structure of the enzyme, which we now call P3H1, demonstrates that P3H1 is a member of a family of prolyl 3-hydroxylases, which share the conserved residues present in the active site of prolyl 4-hydroxylase and lysyl hydroxylase. P3H1 is the chick homologue of mammalian leprecan or growth suppressor 1. Two other P3H family members are the genes previously called MLAT4 and GRCR In this study we demonstrate prolyl 3-hydroxylase activity of the purified enzyme P3H1 on a full-length procollagen substrate. We also show it to specifically interact with denatured collagen and to exist in a tight complex with other endoplasmic reticulum-resident proteins. Immunohistochemistry with a monoclonal antibody specific for chick P3H1 localizes P3H1 specifically to tissues that express fibrillar collagens, suggesting that other P3H family members may be responsible for modifying basement membrane collagens.",
author = "Janice Vranka and Lynn Sakai and B{\"a}chinger, {Hans Peter}",
year = "2004",
month = "5",
day = "28",
doi = "10.1074/jbc.M312807200",
language = "English (US)",
volume = "279",
pages = "23615--23621",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "22",

}

TY - JOUR

T1 - Prolyl 3-hydroxylase 1, enzyme characterization and identification of a novel family of enzymes

AU - Vranka, Janice

AU - Sakai, Lynn

AU - Bächinger, Hans Peter

PY - 2004/5/28

Y1 - 2004/5/28

N2 - The collagen prolyl hydroxylases are enzymes that are required for proper collagen biosynthesis, folding, and assembly. They reside within the endoplasmic reticulum and belong to the group of 2-oxoglutarate and iron-dependent dioxygenases. Although prolyl 4-hydroxylase has been characterized as an α2β2 tetramer in which protein disulfide isomerase is the β subunit with two different α subunit isoforms, little is known about the enzyme prolyl 3-hydroxylase (P3H). It was initially characterized and shown to have an enzymatic activity distinct from that of prolyl 4-hydroxylase, but no amino acid sequences or genes were ever reported for the mammalian enzyme. Here we report the characterization of a novel prolyl 3-hydroxylase enzyme isolated from embryonic chicks. The primary structure of the enzyme, which we now call P3H1, demonstrates that P3H1 is a member of a family of prolyl 3-hydroxylases, which share the conserved residues present in the active site of prolyl 4-hydroxylase and lysyl hydroxylase. P3H1 is the chick homologue of mammalian leprecan or growth suppressor 1. Two other P3H family members are the genes previously called MLAT4 and GRCR In this study we demonstrate prolyl 3-hydroxylase activity of the purified enzyme P3H1 on a full-length procollagen substrate. We also show it to specifically interact with denatured collagen and to exist in a tight complex with other endoplasmic reticulum-resident proteins. Immunohistochemistry with a monoclonal antibody specific for chick P3H1 localizes P3H1 specifically to tissues that express fibrillar collagens, suggesting that other P3H family members may be responsible for modifying basement membrane collagens.

AB - The collagen prolyl hydroxylases are enzymes that are required for proper collagen biosynthesis, folding, and assembly. They reside within the endoplasmic reticulum and belong to the group of 2-oxoglutarate and iron-dependent dioxygenases. Although prolyl 4-hydroxylase has been characterized as an α2β2 tetramer in which protein disulfide isomerase is the β subunit with two different α subunit isoforms, little is known about the enzyme prolyl 3-hydroxylase (P3H). It was initially characterized and shown to have an enzymatic activity distinct from that of prolyl 4-hydroxylase, but no amino acid sequences or genes were ever reported for the mammalian enzyme. Here we report the characterization of a novel prolyl 3-hydroxylase enzyme isolated from embryonic chicks. The primary structure of the enzyme, which we now call P3H1, demonstrates that P3H1 is a member of a family of prolyl 3-hydroxylases, which share the conserved residues present in the active site of prolyl 4-hydroxylase and lysyl hydroxylase. P3H1 is the chick homologue of mammalian leprecan or growth suppressor 1. Two other P3H family members are the genes previously called MLAT4 and GRCR In this study we demonstrate prolyl 3-hydroxylase activity of the purified enzyme P3H1 on a full-length procollagen substrate. We also show it to specifically interact with denatured collagen and to exist in a tight complex with other endoplasmic reticulum-resident proteins. Immunohistochemistry with a monoclonal antibody specific for chick P3H1 localizes P3H1 specifically to tissues that express fibrillar collagens, suggesting that other P3H family members may be responsible for modifying basement membrane collagens.

UR - http://www.scopus.com/inward/record.url?scp=2542497037&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2542497037&partnerID=8YFLogxK

U2 - 10.1074/jbc.M312807200

DO - 10.1074/jbc.M312807200

M3 - Article

C2 - 15044469

AN - SCOPUS:2542497037

VL - 279

SP - 23615

EP - 23621

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 22

ER -