Research output per year
Research output per year
Alexander Eckersley, Kieran T Mellody, Suzanne M Pilkington, Christopher Em Griffiths, Rachel E B Watson, Ronan O'Cualain, Clair Baldock, David Knight, Michael J Sherratt
Research output: Contribution to journal › Article › peer-review
Elastic fibres comprising fibrillin microfibrils and elastin are present in many tissues, including the skin, lung, and arteries where they confer elasticity and resilience. Although fibrillin microfibrils play distinct and tissue-specific functional roles, it is unclear whether their ultrastructure and composition differ between elastin-rich (skin) and elastin-poor (ciliary body and zonule) organs or after in vitro synthesis by cultured cells. Here, we used atomic force microscopy, which revealed that the bead morphology of fibrillin microfibrils isolated from the human eye differs from those isolated from the skin. Using newly developed pre-MS preparation methods and LC-MS/MS, we detected tissue-specific regions of the fibrillin-1 primary structure that were differentially susceptible to proteolytic extraction. Comparing tissue- and culture-derived microfibrils, we found that dermis- and dermal fibroblast-derived fibrillin microfibrils differ in both bead morphology and periodicity and also exhibit regional differences in fibrillin-1 proteolytic susceptibility. In contrast, collagen VI microfibrils from the same dermal or fibroblast samples were invariant in ultrastructure (periodicity) and protease susceptibility. Finally, we observed that skin- and eye-derived microfibril suspensions were enriched in elastic fibre- and basement membrane-associated proteins, respectively. LC-MS/MS also identified proteins (such as calreticulin and protein disulphide isomerase) that are potentially fundamental to fibrillin microfibril biology, regardless of their tissue source. Fibrillin microfibrils synthesised in cell culture lacked some of these key proteins (MFAPs 2 and 4 and fibrillin-2). These results showcase the structural diversity of these key extracellular matrix assemblies, which may relate to their distinct roles in the tissues where they reside.
Original language | English |
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Pages (from-to) | 5117-5133 |
Number of pages | 17 |
Journal | The Journal of biological chemistry |
Volume | 293 |
Issue number | 14 |
Early online date | 16 Feb 2018 |
DOIs | |
Publication status | Published - 6 Apr 2018 |
Research output: Chapter in Book/Conference proceeding › Conference contribution
Eckersley, A. (Owner), Mellody, K. T. (Contributor), Pilkington, S. (Contributor), Griffiths, C. (Contributor), Watson, R. (Supervisor), O’Cualain, R. (Data Collector), Baldock, C. (Contributor), Knight, D. (Data Collector) & Sherratt, M. (Supervisor), ProteomeXchange, 16 Feb 2018
DOI: 10.6019/PXD008450, https://www.ebi.ac.uk/pride/archive/projects/PXD008450 and one more link, https://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD008450 (show fewer)
Dataset
Knight, D. (Platform Lead), Warwood, S. (Senior Technical Specialist), Selley, J. (Technical Specialist), Taylor, G. (Technical Specialist), Fullwood, P. (Technical Specialist), Keevill, E.-J. (Senior Technician) & Allsey, J. (Technician)
FBMH Platform Sciences, Enabling Technologies & InfrastructureFacility/equipment: Facility