A comprehensive map of human elastin cross-linking during elastogenesis.
Elastin is an essential structural protein in the extracellular matrix of vertebrates. It is the core component of elastic fibers, which enable connective tissues such as those of the skin, lungs or blood vessels to stretch and recoil. This function is provided by elastin's exceptional properties, which mainly derive from a unique covalent cross-linking between hydrophilic lysine-rich motifs of units of the monomeric precursor tropoelastin. To date, elastin's cross-linking is poorly investigated. Here, we purified elastin from human tissue and cleaved it into soluble peptides using proteases with different specificities. We then analyzed elastin's molecular structure by identifying unmodified residues, post-translational modifications and cross-linked peptides by high-resolution mass spectrometry and amino acid analysis. The data revealed the presence of multiple isoforms in parallel and a complex and heterogeneous molecular interconnection. We discovered that the same lysine residues in different monomers were simultaneously involved in various cross-link types or remained unmodified. Furthermore, both types of cross-linking domains, Lys-Pro and Lys-Ala domains, participate not only in bifunctional inter- but also in intra-domain cross-links. We elucidated the sequences of several desmosine-containing peptides and the contribution of distinct domains such as 6, 14 and 25. In contrast to earlier assumptions proposing that desmosine cross-links are formed solely between two domains, we elucidated the structure of a peptide that proves a desmosine formation with participation of three Lys-Ala domains. In summary, these results provide new and detailed insights into the cross-linking process, which takes place within and between human tropoelastin units in a stochastic manner.
['Hedtke|Tobias|T|0000-0002-2662-6576', 'Schräder|Christoph U|CU|0000-0002-0461-3782', 'Heinz|Andrea|A|0000-0002-8609-4460', 'Hoehenwarter|Wolfgang|W|0000-0002-7669-7524', 'Brinckmann|Jürgen|J|', 'Groth|Thomas|T|0000-0001-6647-9657', 'Schmelzer|Christian E H|CEH|0000-0002-1180-0201']
D000595:Amino Acid Sequence D003895:Desmosine D004547:Elastic Tissue D004549:Elastin D005109:Extracellular Matrix D006801:Humans D057927:Hydrophobic and Hydrophilic Interactions D008239:Lysine D013058:Mass Spectrometry D015394:Molecular Structure D010455:Peptides D020033:Protein Isoforms D011499:Protein Processing, Post-Translational D012867:Skin D014333:Tropoelastin
extracellular matrix elastic fibers mass spectrometry desmosine lysyl oxidase