Extracellular vesicles and post-translational protein deimination signatures in haemolymph of the American lobster (Homarus americanus)

Post-Translational Protein Deimination Signatures in Plasma and Plasma EVs of Reindeer (Rangifer tarandus)

“…In whole plasma, key complement components including C1q, C3, C4, C5, C9, as well as factor H and the C3/C5 convertase were identified as deiminated. Deimination of complement components has previously been reported for a various complement components, including some identified here, in serum and plasma from a range of species [12][13][14][15][16][17][18][19][20][21][22][23][24][25]29]. Interestingly, in bovine serum-EVs, a number of deiminated complement components has been identified, including C1q, C3, C4A, C5a, C7, C8, C9 factor B, Factor H, C4-binding protein [20], while in teleost fish serum both C3 and C4 were identified in deiminated form but in teleost serum-EVs C3 was more dominant in deiminated form, compared with C4 [12,13,16].…”

“…Primary antibody incubation was carried out overnight at 4 • C on a shaking platform using the following antibodies for reindeer plasma: F95 pan-deimination antibody (MABN328, Merck; diluted 1/1000 in TBS-T) and anti-human PAD2 (ab50257, Abcam, diluted 1/1000 in TBS-T), PAD3 (ab50246, diluted 1/1000 in TBS-T) and PAD4 (ab50332, diluted 1/1000 in TBS-T) antibodies, for detection of putative PAD protein homologues. PAD2 is considered the most conserved PAD isozyme and the anti-human PAD2 antibody was previously shown to cross-react with PADs across taxa [17][18][19][20][21][22][23][24][25]28,29,60,61], while both the PAD3 and PAD4 antibodies have also been found to cross react with other species, including bird, reptile and bovine [18,20,60]. EV-cargo was also assessed for PAD2, PAD3 and PAD4 as well as deiminated proteins (F95).…”

“…While the reindeer genome has been sequenced [10], and genetic diversity and mitochondrial DNA have furthermore been studied [11], no studies have hitherto been performed into mechanisms relating to post-translational modifications such as deimination, which is caused by peptidylarginine deiminases (PADs). Furthermore, while research on extracellular vesicles (EVs) is a major field in relation to biomarker discovery in human pathologies, and recent comparatives studies have highlighted their value in a range of wild, domestic and commercially valuable land and aquatic animals throughout the phylogeny tree [12][13][14][15][16][17][18][19][20][21][22][23][24][25], the field is still in its infancy in relation to studies and biomarker development in wild animals.…”

“…A majority of studies on PADs and downstream deimination have related to human pathological mechanisms, but recently a comparative body of research has focused on identifying putative roles for PADs in physiological and immunological pathways in a wide range of taxa throughout the phylogenetic tree, including land and sea mammals, reptiles, birds, bony and cartilaginous fish, Mollusca and Crustacea [16][17][18][19][20][21][22][23][24][25]28,29]. PADs have furthermore been identified to have roles in mucosal, innate and adaptive immunity in a range of taxa [17][18][19][20]25,28,29,43,44].…”

Extracellular Vesicles and Post-Translational Protein Deimination Signatures in Mollusca—The Blue Mussel (Mytilus edulis), Soft Shell Clam (Mya arenaria), Eastern Oyster (Crassostrea virginica) and Atlantic Jacknife Clam (Ensis leei)

“…A majority of studies on PADs and downstream deimination have hitherto related to human pathological mechanisms, but recently a comparative body of research has focused on identifying putative roles for PADs in physiological and immunological pathways in a wide range of taxa throughout the phylogenetic tree, including land and sea mammals, reptiles, birds, bony, and cartilaginous fish, Myrostomata and Crustacea. In these studies, PADs have indeed been identified to have roles in mucosal, innate, and adaptive immunity in a range of taxa [21][22][23][24][25][26][44][45][46][47][48][49][50][51][52][53]. Importantly, PADs have also been identified as important players in infection and antipathogenic responses, including antiviral [54,55], antiparasitic [28], and antibacterial ones [29,30].…”