Comprehensive mass spectrometric mapping of the hydroxylated amino acid residues of the α1(V) collagen chain

Chenxi Yang, Arick C. Park, Nicholas A. Davis, Jason D. Russell, Byoungjae Kim, David Brand, Matthew J. Lawrence, Ying Ge, Michael S. Westphall, Joshua J. Coon, Daniel S. Greenspan

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Aberrant expression of the type V collagen α1(V) chain can underlie the connective tissue disorder classic Ehlers-Danlos syndrome, and autoimmune responses against the α1(V) chain are linked to lung transplant rejection and atherosclerosis. The α1(V) collagenous COL1 domain is thought to contain greater numbers of post-translational modifications (PTMs) than do similar domains of other fibrillar collagen chains, PTMs consisting of hydroxylated prolines and lysines, the latter of which can be glycosylated. These types of PTMs can contribute to epitopes that underlie immune responses against collagens, and the high level of PTMs may contribute to the unique biological properties of the α1(V) chain. Here we use high resolution mass spectrometry to map such PTMs in bovine placental α1(V) and human recombinant pro-α1(V) procollagen chains. Findings include the locations of those PTMs that vary and those PTMs that are invariant between these α1(V) chains from widely divergent sources. Notably, an unexpectedly large number of hydroxyproline residues were mapped to the X-positions of Gly-X-Y triplets, contrary to expectations based on previous amino acid analyses of hydrolyzed α1(V) chains from various tissues. We attribute this difference to the ability of tandem mass spectrometry coupled to nanoflow chromatographic separations to detect lower-level PTM combinations with superior sensitivity and specificity. The data are consistent with the presence of a relatively large number of 3-hydroxyproline sites with less than 100% occupancy, suggesting a previously unknown mechanism for the differential modification of α1(V) chain and type V collagen properties.

Original languageEnglish (US)
Pages (from-to)40598-40610
Number of pages13
JournalJournal of Biological Chemistry
Volume287
Issue number48
DOIs
StatePublished - Nov 23 2012

Fingerprint

Collagen Type V
Post Translational Protein Processing
Mass spectrometry
Collagen
Fibrillar Collagens
Tissue
Amino Acids
Procollagen
Transplants
Hydroxyproline
Proline
Lysine
Epitopes
Ehlers-Danlos Syndrome
Graft Rejection
Collagen Type I
Tandem Mass Spectrometry
Autoimmunity
Connective Tissue
Mass Spectrometry

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Comprehensive mass spectrometric mapping of the hydroxylated amino acid residues of the α1(V) collagen chain. / Yang, Chenxi; Park, Arick C.; Davis, Nicholas A.; Russell, Jason D.; Kim, Byoungjae; Brand, David; Lawrence, Matthew J.; Ge, Ying; Westphall, Michael S.; Coon, Joshua J.; Greenspan, Daniel S.

In: Journal of Biological Chemistry, Vol. 287, No. 48, 23.11.2012, p. 40598-40610.

Research output: Contribution to journalArticle

Yang, C, Park, AC, Davis, NA, Russell, JD, Kim, B, Brand, D, Lawrence, MJ, Ge, Y, Westphall, MS, Coon, JJ & Greenspan, DS 2012, 'Comprehensive mass spectrometric mapping of the hydroxylated amino acid residues of the α1(V) collagen chain', Journal of Biological Chemistry, vol. 287, no. 48, pp. 40598-40610. https://doi.org/10.1074/jbc.M112.406850
Yang, Chenxi ; Park, Arick C. ; Davis, Nicholas A. ; Russell, Jason D. ; Kim, Byoungjae ; Brand, David ; Lawrence, Matthew J. ; Ge, Ying ; Westphall, Michael S. ; Coon, Joshua J. ; Greenspan, Daniel S. / Comprehensive mass spectrometric mapping of the hydroxylated amino acid residues of the α1(V) collagen chain. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 48. pp. 40598-40610.
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AU - Kim, Byoungjae

AU - Brand, David

AU - Lawrence, Matthew J.

AU - Ge, Ying

AU - Westphall, Michael S.

AU - Coon, Joshua J.

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AB - Aberrant expression of the type V collagen α1(V) chain can underlie the connective tissue disorder classic Ehlers-Danlos syndrome, and autoimmune responses against the α1(V) chain are linked to lung transplant rejection and atherosclerosis. The α1(V) collagenous COL1 domain is thought to contain greater numbers of post-translational modifications (PTMs) than do similar domains of other fibrillar collagen chains, PTMs consisting of hydroxylated prolines and lysines, the latter of which can be glycosylated. These types of PTMs can contribute to epitopes that underlie immune responses against collagens, and the high level of PTMs may contribute to the unique biological properties of the α1(V) chain. Here we use high resolution mass spectrometry to map such PTMs in bovine placental α1(V) and human recombinant pro-α1(V) procollagen chains. Findings include the locations of those PTMs that vary and those PTMs that are invariant between these α1(V) chains from widely divergent sources. Notably, an unexpectedly large number of hydroxyproline residues were mapped to the X-positions of Gly-X-Y triplets, contrary to expectations based on previous amino acid analyses of hydrolyzed α1(V) chains from various tissues. We attribute this difference to the ability of tandem mass spectrometry coupled to nanoflow chromatographic separations to detect lower-level PTM combinations with superior sensitivity and specificity. The data are consistent with the presence of a relatively large number of 3-hydroxyproline sites with less than 100% occupancy, suggesting a previously unknown mechanism for the differential modification of α1(V) chain and type V collagen properties.

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