Crystallization of the proline-rich-peptide binding domain of human type I collagen prolyl 4-hydroxylase

Pekkala, M.; Hieta, Reija; Kursula, Petri; Kivirikko, Kari I.; Wierenga, Rik K.; Myllyharju, Johanna

doi:10.1107/s0907444903005420

Cited by 14 publications

(15 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Many of the characteristic properties found in the binding of peptide substrates and inhibitors to the type I and type II C-P4Hs can be explained by the properties of this domain rather than by those of the catalytic domain. We have recently obtained crystals of this domain that diffract to at least 3 Å resolution (40), and attempts to determine the complete crystal-based structure are now in progress. values, binding stoichiometry (n), and change in enthalpy (⌬H) of the ␣(I) peptide binding domain, determined by isothermal titration calorimetry The data were measured using 50-500 M of the peptide binding domain and 10-20-fold molar excesses of the different peptides at 25°C and fitted using a nonlinear least squares algorithm.…”

Section: Discussionmentioning

confidence: 99%

The Peptide-Substrate-binding Domain of Human Collagen Prolyl 4-Hydroxylases

Hieta

Kukkola

Permi

et al. 2003

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The collagen prolyl 4-hydroxylases (C-P4Hs) catalyze the formation of 4-hydroxyproline by the hydroxylation of proline residues in -Xaa-Pro-Gly-sequences. The vertebrate enzymes are ␣ 2 ␤ 2 tetramers in which protein-disulfide isomerase serves as the ␤ subunit. Two isoforms of the catalytic ␣ subunit have been identified and shown to form [␣(I)] 2 ␤ 2 and [␣(II)] 2 ␤ 2 tetramers, the type I and type II C-P4Hs, respectively. The peptide-substrate-binding domain of type I C-P4H has been shown to be located between residues 138 and 244 in the 517-residue ␣(I) subunit and to be distinct from the catalytic domain that is located in the C-terminal region. We report here that a recombinant human C-P4H ␣(I) polypeptide The prolyl 4-hydroxylases (P4Hs) 1 catalyze the formation of 4-hydroxyproline by the hydroxylation of proline residues in peptide linkages. Two P4H families are known today. The collagen P4Hs (C-P4Hs), enzymes residing within the lumen of the endoplasmic reticulum, have a central role in the synthesis of all collagens, the resulting 4-hydroxyproline residues being essential for assembly of the triple-helical molecules (1-3). The hypoxia-inducible factor (HIF) P4Hs, a family of cytoplasmic enzymes (4 -6), play a key role in the response of cells to hypoxia by catalyzing hydroxylation of the ␣ subunit of HIF. This subunit is synthesized continuously, and at least one of two critical proline residues becomes hydroxylated under normoxic conditions, the resulting 4-hydroxyproline being essential for rapid degradation of HIF-␣ (7-9). In hypoxia this hydroxylation ceases, HIF-␣ forms a dimer with HIF-␤, and the dimer then becomes bound to the HIF-responsive elements in a number of hypoxia-inducible genes.All vertebrate C-P4Hs are ␣ 2 ␤ 2 tetramers in which the ␤ subunit is identical to the enzyme and chaperone protein-disulfide isomerase (1-3). Two isoforms of the catalytic ␣ subunit have been characterized from human and mouse tissues and shown to form [␣(I)] 2 ␤ 2 and [␣(II)] 2 ␤ 2 tetramers, called the type I and type II C-P4Hs, respectively (10, 11). The type I enzyme is the most abundant form in most cells, but type II is the main form in chondrocytes, endothelial cells, and some other cell types (12, 13). The HIF-P4Hs appear to consist of only one type of monomer, the size of which ranges from 239 to 426 residues in the three human isoenzymes (4 -6).The C-P4Hs act on -Xaa-Pro-Gly-triplets in collagens and more than 15 other proteins with collagen-like sequences (1-3, 14), whereas the HIF-P4Hs hydroxylate -Leu-Xaa-Xaa-LeuAla-Pro-Tyr-and -Leu-Xaa-Xaa-Leu-Ala-Pro-Ala-sequences (7-9). All the P4Hs require Fe 2ϩ , 2-oxoglutarate, O 2 , and ascorbate, the 2-oxoglutarate being stoichiometrically decarboxylated during hydroxylation (1-3). The C-terminal regions of the C-P4H ␣ subunits and the HIF-P4H monomers contain four conserved residues, two histidines, and one aspartate that bind the Fe 2ϩ atom and a basic residue that binds the C-5 carboxyl group of the 2-oxoglutarate (15,16). This basic residue is a lysine ...

show abstract

Section: Discussionmentioning

confidence: 99%

The Peptide-Substrate-binding Domain of Human Collagen Prolyl 4-Hydroxylases

Hieta

Kukkola

Permi

et al. 2003

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…The most extensively studied prolyl hydroxylases are the ones that hydroxylate a proline residues in collagen molecules (8)(9)(10)(11)(12)(13). These studies reveal that the enzyme, isolated as a homogeneous protein by affinity chromatography from three different sources (8)(9)(10)12,14,15), occurs as a tetramer with a molecular weight of about 240,000 (11,(16)(17)(18). The enzyme does not hydroxylate free proline, and recognizes a conserved motif (LXXLAP in the HIF-1 molecule; X indicates any amino acid and P indicates the hydroxyl acceptor proline) in the primary substrate for hydroxylation (5,11,(19)(20)(21)(22).…”

Section: Regulation Of Prolyl 4-hydroxylase (Phd) Enzyme Activity Viamentioning

confidence: 99%

Prolyl 4-hydroxylase activity-responsive transcription factors: From hydroxylation to gene expression and neuroprotection

Siddiq

Aminova²,

Ratan³

2008

Front Biosci

View full text Add to dashboard Cite

Most homeostatic processes including gene transcription occur as a result of deviations in physiological tone that threatens the survival of the organism. A prototypical homeostatic stress response includes changes in gene expression following alterations in oxygen, iron or 2-oxoglutarate levels. Each of these cofactors plays an important role in cellular metabolism. Accordingly, a family of enzymes known as the Prolyl 4-hydroxylase (PHD) enzymes are a group of dioxygenases that have evolved to sense changes in 2-oxoglutarate, oxygen and iron via changes in enzyme activity. Indeed, PHDs are a part of an established oxygen sensor system that regulates transcriptional regulation of hypoxia/stress-regulated genes and thus are an important component of events leading to cellular rescue from oxygen, iron or 2-oxoglutarate deprivations. The ability of PHD activity to regulate homeostatic responses to oxygen, iron or 2-oxoglutarate metabolism has led to the development of small molecule inhibitors of the PHDs as a strategy for activating or augmenting cellular stress responses. These small molecules are proving effective in preclinical models of stroke and Parkinson's disease. However the precise protective pathways engaged by PHD inhibition are only beginning to be defined. In the current review, we summarize the role of iron, 2-oxoglutarate and oxygen in the PHD catalyzed hydroxylation reaction and provide a brief discussion of some of the transcription factors that play an effective role in neuroprotection against oxidative stress as a result of changes in PHD activity.

show abstract

“…We recently obtained crystals of the ␣(I) peptide-substratebinding domain (23), and we have now obtained crystals that diffract to 2.3 Å resolution. The domain was found to consist of five ␣-helices and belong to the family of tetratricopeptide repeat (TPR) domains that are involved in many protein-protein interactions (24 -27).…”

mentioning

confidence: 99%

“…The recombinant domain was expressed in Escherichia coli and purified as described earlier (14,23). Se-Met derivatization of the protein was performed by expressing it in a methionine-requiring auxotroph E. coli strain B384 (DE3) using induction for 20 h at 20°C.…”

mentioning

confidence: 99%

See 1 more Smart Citation

The Peptide-Substrate-binding Domain of Collagen Prolyl 4-Hydroxylases Is a Tetratricopeptide Repeat Domain with Functional Aromatic Residues

Pekkala¹,

Hieta

Bergmann³

et al. 2004

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Collagen prolyl 4-hydroxylases catalyze the formation of 4-hydroxyproline in -X-Pro-Gly-sequences and have an essential role in collagen synthesis. The vertebrate enzymes are ␣ 2 ␤ 2 tetramers in which the catalytic ␣-subunits contain separate peptide-substrate-binding and catalytic domains. We report on the crystal structure of the peptide-substrate-binding domain of the human type I enzyme refined at 2.3 Å resolution. It was found to belong to a family of tetratricopeptide repeat domains that are involved in many protein-protein interactions and consist of five ␣-helices forming two tetratricopeptide repeat motifs plus the solvating helix. A prominent feature of its concave surface is a deep groove lined by tyrosines, a putative binding site for proline-rich peptides. Solvent-exposed side chains of three of the tyrosines have a repeat distance similar to that of a poly-L-proline type II helix. The aromatic surface ends at one of the tyrosines, where the groove curves almost 90°a way from the linear arrangement of the three tyrosine side chains, possibly inducing a bent conformation in the bound peptide. This finding is consistent with previous suggestions by others that a minimal structural requirement for proline 4-hydroxylation may be a sequence in the poly-L-proline type II conformation followed by a ␤-turn in the Pro-Gly segment. Site-directed mutagenesis indicated that none of the tyrosines was critical for tetramer assembly, whereas most of them were critical for the binding of a peptide substrate and inhibitor both to the domain and the ␣ 2 ␤ 2 enzyme tetramer.

show abstract

Crystallization of the proline-rich-peptide binding domain of human type I collagen prolyl 4-hydroxylase

Cited by 14 publications

References 17 publications

The Peptide-Substrate-binding Domain of Human Collagen Prolyl 4-Hydroxylases

The Peptide-Substrate-binding Domain of Human Collagen Prolyl 4-Hydroxylases

Prolyl 4-hydroxylase activity-responsive transcription factors: From hydroxylation to gene expression and neuroprotection

The Peptide-Substrate-binding Domain of Collagen Prolyl 4-Hydroxylases Is a Tetratricopeptide Repeat Domain with Functional Aromatic Residues

Contact Info

Product

Resources

About