Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
PLoS One
2014 Jan 01;94:e93467. doi: 10.1371/journal.pone.0093467.
Show Gene links
Show Anatomy links
Evolutionary origins of C-terminal (GPP)n 3-hydroxyproline formation in vertebrate tendon collagen.
Hudson DM
,
Werther R
,
Weis M
,
Wu JJ
,
Eyre DR
.
???displayArticle.abstract???
Approximately half the proline residues in fibrillar collagen are hydroxylated. The predominant form is 4-hydroxyproline, which helps fold and stabilize the triple helix. A minor form, 3-hydroxyproline, still has no clear function. Using peptide mass spectrometry, we recently revealed several previously unknown molecular sites of 3-hydroxyproline in fibrillar collagen chains. In fibril-forming A-clade collagen chains, four new partially occupied 3-hydroxyproline sites were found (A2, A3, A4 and (GPP)n) in addition to the fully occupied A1 site at Pro986. The C-terminal (GPP)n motif has five consecutive GPP triplets in α1(I), four in α2(I) and three in α1(II), all subject to 3-hydroxylation. The evolutionary origins of this substrate sequence were investigated by surveying the pattern of its 3-hydroxyproline occupancy from early chordates through amphibians, birds and mammals. Different tissue sources of type I collagen (tendon, bone and skin) and type II collagen (cartilage and notochord) were examined by mass spectrometry. The (GPP)n domain was found to be a major substrate for 3-hydroxylation only in vertebrate fibrillar collagens. In higher vertebrates (mouse, bovine and human), up to five 3-hydroxyproline residues per (GPP)n motif were found in α1(I) and four in α2(I), with an average of two residues per chain. In vertebrate type I collagen the modification exhibited clear tissue specificity, with 3-hydroxyproline prominent only in tendon. The occupancy also showed developmental changes in Achilles tendon, with increasing 3-hydroxyproline levels with age. The biological significance is unclear but the level of 3-hydroxylation at the (GPP)n site appears to have increased as tendons evolved and shows both tendon type and developmental variations within a species.
???displayArticle.pubmedLink???
24695516
???displayArticle.pmcLink???PMC3973637 ???displayArticle.link???PLoS One ???displayArticle.grants???[+]
Figure 2. Mass spectra of (GPP)n containing tryptic peptides from adult animal tendons.Full scan spectra from LC-MS profiles of in-gel trypsin digests of α1(I) from human, chicken and xenopus tendon with 6% SDS-PAGE lanes at left (A). MS/MS fragmentation spectrum of the parent ion (1265.33+) from human tendon (B). The sequence is shown with b and y ion breakages. P*, 4Hyp; P#, 3Hyp; K*, Hyl. The cross-linking telopeptide Lys of the human peptide was fully hydroxylated in all posttranslational variants.
Figure 3. Edman N-terminal sequence analysis confirming 3-hydroxyproline in the (GPP)n motif of tendon type I collagen.The (GPP)n-containing tryptic peptides from the α1(I) and α2(I) α-chains of rat tail tendon were recovered as a chromatographic pool and sequenced simultaneously. Sequential phenylthiohydantoin-derivative HPLC chromatograms are shown for sequencer cycles 8-15 (sequencer cycles 1-7 were as predicted from the known sequences of both α-chains). The new residues at each cycle are highlighted in blue for the α1-chain and green for the α2-chain. The 3Hyp residue gave a distinctive double peak as previously reported [18].
Figure 4. Mass spectra showing prolyl 3-hydroxylation distributed throughout the whole fibril.Collagen was solubilized from adult human tendon using SDS extraction (A) and CNBr digestion (B). Lanes of 6% (A) and 12% (B) SDS-PAGE gels are shown to the left. Similar levels of 3Hyp (∼two 3Hyp per α2(I) chain) were observed using each approach. The 12102+ and 12182+ ions in both ion ladders represent unrelated peptides with a 2+ charge (these ions are indicated with φ).
Figure 5. Developmental control of prolyl 3-hydroxylation in tendon.Reduced levels of 3Hyp were observed in fetal tendon relative to adult tissue. MS scan of fetal human Achilles tendon α1(I) with 6% SDS-PAGE lane at left (A). The 1248.43+ ion contains a mix of two peptide posttranslational variants (one 3Hyp and four 4Hyp; and no 3Hyp and five 4Hyp). MS scan of fetal human Achilles tendon α2(I) (B).
Figure 1. Protein sequence alignment of the collagen (GPP)n motif from phylogenetically diverse animals.Conservation of the (GPP)n motif is shown in red for fibrillar collagens from early chordates through amphibians, birds and mammals. Genomic sequences are from Ensembl. Lamprey and ciona collagen sequences are from Petromyzon marinus transcript: COL2A1 ENSPMAT00000009617 and Ciona intestinalis transcript: FCOL1 ENSCINT00000014311, respectively.
Bailey,
Molecular mechanisms of ageing in connective tissues.
2001, Pubmed
Bailey,
Molecular mechanisms of ageing in connective tissues.
2001,
Pubmed
Brondijk,
Implications for collagen I chain registry from the structure of the collagen von Willebrand factor A3 domain complex.
2012,
Pubmed
Byers,
Recessively inherited forms of osteogenesis imperfecta.
2012,
Pubmed
Cabral,
Prolyl 3-hydroxylase 1 deficiency causes a recessive metabolic bone disorder resembling lethal/severe osteogenesis imperfecta.
2007,
Pubmed
Canty,
Coalignment of plasma membrane channels and protrusions (fibripositors) specifies the parallelism of tendon.
2004,
Pubmed
Capellini,
Conservation of notochord gene expression across chordates: insights from the Leprecan gene family.
2008,
Pubmed
Cundy,
Recent advances in osteogenesis imperfecta.
2012,
Pubmed
Ehrlich,
Mineralization of the metre-long biosilica structures of glass sponges is templated on hydroxylated collagen.
2010,
Pubmed
Eyre,
Cross-linking in collagen and elastin.
1984,
Pubmed
Eyre,
A novel 3-hydroxyproline (3Hyp)-rich motif marks the triple-helical C terminus of tendon type I collagen.
2011,
Pubmed
Fernandes,
A role for prolyl 3-hydroxylase 2 in post-translational modification of fibril-forming collagens.
2011,
Pubmed
Ha-Vinh,
Phenotypic and molecular characterization of Bruck syndrome (osteogenesis imperfecta with contractures of the large joints) caused by a recessive mutation in PLOD2.
2004,
Pubmed
Hudson,
Collagen prolyl 3-hydroxylation: a major role for a minor post-translational modification?
2013,
Pubmed
Hudson,
Insights on the evolution of prolyl 3-hydroxylation sites from comparative analysis of chicken and Xenopus fibrillar collagens.
2011,
Pubmed
,
Xenbase
Hudson,
Peptidyl 3-hydroxyproline binding properties of type I collagen suggest a function in fibril supramolecular assembly.
2012,
Pubmed
Kadler,
Collagens at a glance.
2007,
Pubmed
Light,
Changes in crosslinking during aging in bovine tendon collagen.
1979,
Pubmed
Morello,
CRTAP is required for prolyl 3- hydroxylation and mutations cause recessive osteogenesis imperfecta.
2006,
Pubmed
Myllyharju,
Collagens, modifying enzymes and their mutations in humans, flies and worms.
2004,
Pubmed
Parry,
A comparison of the size distribution of collagen fibrils in connective tissues as a function of age and a possible relation between fibril size distribution and mechanical properties.
1978,
Pubmed
Pokidysheva,
Posttranslational modifications in type I collagen from different tissues extracted from wild type and prolyl 3-hydroxylase 1 null mice.
2013,
Pubmed
Pyott,
Mutations in PPIB (cyclophilin B) delay type I procollagen chain association and result in perinatal lethal to moderate osteogenesis imperfecta phenotypes.
2011,
Pubmed
Ricard-Blum,
The collagen family.
2011,
Pubmed
Shadwick,
Elastic energy storage in tendons: mechanical differences related to function and age.
1990,
Pubmed
Shadwick,
Structure and function of tuna tail tendons.
2002,
Pubmed
Steinmann,
Urinary pyridinoline cross-links in Ehlers-Danlos syndrome type VI.
1995,
Pubmed
Summers,
The evolution of tendon--morphology and material properties.
2002,
Pubmed
Tiainen,
Characterization of recombinant human prolyl 3-hydroxylase isoenzyme 2, an enzyme modifying the basement membrane collagen IV.
2008,
Pubmed
Vranka,
Prolyl 3-hydroxylase 1, enzyme characterization and identification of a novel family of enzymes.
2004,
Pubmed
Vranka,
Expression of prolyl 3-hydroxylase genes in embryonic and adult mouse tissues.
2009,
Pubmed
Weis,
Location of 3-hydroxyproline residues in collagen types I, II, III, and V/XI implies a role in fibril supramolecular assembly.
2010,
Pubmed
Widmer,
Molecular basis for the action of the collagen-specific chaperone Hsp47/SERPINH1 and its structure-specific client recognition.
2012,
Pubmed
