Peptide location fingerprinting identifies species- and tissue-conserved structural remodelling of proteins as a consequence of ageing and disease

Extracellular matrices (ECMs) in the intervertebral disc (IVD), lung and artery are thought to undergo age-dependant accumulation of damage by chronic exposure to mechanisms such as reactive oxygen species, proteases and glycation. It is unknown whether this damage accumulation is species-dependant (via differing lifespans and hence cumulative exposures) or whether it can influence the progression of age-related diseases such as atherosclerosis. Peptide location fingerprinting (PLF) is a new proteomic analysis method, capable of the non-targeted identification of structure-associated changes within proteins. Here we applied PLF to publicly available ageing human IVD (outer annulus fibrosus), ageing mouse lung and human arterial atherosclerosis datasets and bioinformatically identified novel target proteins alongside common age-associated differences within protein structures which were conserved between three ECM-rich organs, two species, three IVD tissue regions, sexes and in an age-related disease. We identify peptide yield differences across protein structures which coincide with biological regions, potentially reflecting the functional consequences of ageing or atherosclerosis for macromolecular assemblies (collagen VI), enzyme/inhibitor activity (alpha-2 macroglobulin), activation states (complement C3) and interaction states (laminins, perlecan, fibronectin, filamin-A, collagen XIV and apolipoprotein-B). Furthermore, we show that alpha-2 macroglobulin and collagen XIV exhibit possible shared structural consequences in IVD ageing and arterial atherosclerosis, providing novel links between an age-related disease and intrinsic ageing. Crucially, we also demonstrate that fibronectin, laminin beta chains and filamin-A all exhibit conserved age-associated structural differences between mouse lung and human IVD, providing evidence that ECM, and their associating proteins, may be subjected to potentially similar mechanisms or consequences of ageing across both species

Funding agencies: Manchester Institute for Collaborative Research on Ageing (MICRA); Wal-greens Boots Alliance (WBA) (MJS), UK; the British Heart Foundation Centre ofResearch Excellence (BHF CRE); WellcomeTrust grant (108413/A/15/D); RGC European Union - Hong Kong Research and Innovation Cooperation Co-funding Mechanism (E-HKU703/18); European Union’s Horizon 2020research and innovation program (iPSpine; grant agreement number 825925); Shenz-hen “Key Medical Discipline Construction Fund”(SZXK077); UK Engineering and PhysicalSciences Research Council (EPSRC; EP/V011065/1); Swedish Heart Lung Foundation; Stroke Founda-tion; Olle Engkvist Foundation; Swedish GamlaTjänarinnor Foundation; Linköping University Hospital Research Fund.