Identification of cross-linking pattern in human elastin

• Applicant: Dr. Andrea Heinz
• Subject Area: Pharmacy ; Biochemistry
• Term: from 2010 to 2016
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 188218494

Elastin is an essential protein of the extracellular matrix of vertebrates and possesses unique properties such as elasticity, resilience and extreme durability, which make it crucial for the long-term function of different tissues and organs such as aorta, lung, skin or cartilage. The biopolymer is composed of units of its precursor tropoelastin, which are connected via lysine residues and form a variety of polyfunctional cross-links. Due to its high hydrophobicity and complex cross-linking pattern elastin is completely insoluble and very resistant to enzymatic degradation, which limits structure investigations. Hence, virtually nothing is known about the cross-linking pattern of elastin in different human tissues. It is, however, of major importance to investigate the structure of elastin on the molecular level to better understand its functions in healthy tissues as well as structural changes that go along with cardiovascular diseases such as atherosclerosis, lung emphysema and aortic stenosis. Moreover, detailed insights into the structure of elastic fibers will aid in the development of protein-based biomaterials, which may be used for a variety of medical applications. Therefore, the aim of this project is the development and the application of analytical and bioinformatics methods to characterize the cross-linking pattern of elastin isolated from human aorta. Based on different model systems such as cross-linked elastin peptides, cross-linked elastin polypeptides and cross-linked tropoelastin mass spectrometric methods will be developed, which enable identifying and sequencing cross-links characteristic of elastin. Controlled degradation of isolated elastin using specific elastases and subsequent identification of cross-linked peptides will then allow to determine the exact positions of inter- and intramolecular cross-links as well as the domains of tropoelastin molecules involved in cross-linking. The project will provide a comprehensive and significant insight into the structure of elastin. Overall, understanding the structure of elastin will support the development of directed therapies against diseases of elastic tissues, including enzyme inhibitors, wound regeneration materials or vascular prostheses.

• DFG Programme: Research Grants