Analysis of collagen and elastin cross-links

Methods Cell Biol. 2018:143:115-132. doi: 10.1016/bs.mcb.2017.08.006. Epub 2017 Nov 22.

Abstract

Fibrillar collagens represent the most abundant extracellular matrix proteins in vertebrates providing tissues and organs with form, stability, and connectivity. For such mechanical functions, the formation of covalent intermolecular cross-linking between molecules is essential. This process, the final posttranslational modification during collagen biosynthesis, is initiated by conversion of specific lysine and hydroxylysine residues to the respective aldehydes by the action of lysyl oxidases. This conversion triggers a series of condensation reactions with the juxtaposed lysine-aldehyde, lysine, hydroxylysine, and histidine residues within the same and neighboring molecules resulting in di-, tri-, and tetravalent cross-links. Elastin, another class of extracellular matrix protein, is also stabilized by the lysyl oxidase-mediated mechanism but involving only lysine residues leading to the formation of unique tetravalent cross-links. This chapter presents an overview of fibrillar collagen cross-linking, and the analytical methods for collagen and elastin cross-links we have developed.

Keywords: Collagen cross-links; Elastin cross-links; Glycosylation of cross-links; HPLC analysis; Mass spectrometric analysis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Cross-Linking Reagents / chemistry*
  • Elastin / chemistry*
  • Extracellular Matrix / metabolism*
  • Fibrillar Collagens / biosynthesis
  • Fibrillar Collagens / chemistry*
  • Hydrolysis
  • Hydroxylysine / metabolism
  • Lysine / metabolism
  • Protein Processing, Post-Translational / physiology
  • Protein-Lysine 6-Oxidase / metabolism

Substances

  • Cross-Linking Reagents
  • Fibrillar Collagens
  • Hydroxylysine
  • Elastin
  • Protein-Lysine 6-Oxidase
  • Lysine