Abstract
BackgroundSalmon trypsin is shown to increase secretion of the pro-inflammatory cytokine interleukin (IL)-8 from human airway epithelial cells through activation of PAR-2. Secretion of IL-8 induced by king crab trypsin is observed in a different concentration range compared to salmon trypsin, and seems to be only partially related to PAR-2 activation. This report aim to identify differences in the molecular structure of king crab trypsin (Paralithodes camtschaticus) compared to salmon (Salmo salar) and bovine trypsin (Bos taurus) that might influence the ability to activate protease-activated receptor-2 (PAR-2).ResultsDuring purification king crab trypsin displayed stronger binding capacity to the anionic column used in fast protein liquid chromatography compared to fish trypsins, and was identified as a slightly bigger molecule. Measurements of enzymatic activity yielded no obvious differences between the trypsins tested. Molecular modelling showed that king crab trypsin has a large area with strong negative electrostatic potential compared to the smaller negative areas in bovine and salmon trypsins. Bovine and salmon trypsins also displayed areas with strong positive electrostatic potential, a feature lacking in the king crab trypsin. Furthermore we have identified 3 divergent positions (Asp196, Arg244, and Tyr247) located near the substrate binding pocket of king crab trypsin that might affect the binding and cleavage of PAR-2.ConclusionThese preliminary results indicate that electrostatic interactions could be of importance in binding, cleavage and subsequent activation of PAR-2.
Highlights
Salmon trypsin is shown to increase secretion of the pro-inflammatory cytokine interleukin (IL)-8 from human airway epithelial cells through activation of protease-activated receptor-2 (PAR-2)
Our previous work has confirmed that purified salmon trypsin increase secretion of the pro-inflammatory cytokine interleukin (IL)-8 from human airway epithelial cells through activation of PAR-2 [6]
During purification of numerous fish trypsins (Atlantic salmon [Salmo salar], sardine [Sardinops melanostictus], anchovy [Engraulis japonicus], jacopever [Sebastes schlegelii], yellow tail [Seriola quinqueradiata], spotted mackerel [Scomber australasicus] ) and trypsin from the king crab (Paralithodes camtschaticus) by fast protein liquid chromatography (FPLC), we observed that king crab trypsin bound stronger to the anionic column compared to the fish trypsins we purified
Summary
Salmon trypsin is shown to increase secretion of the pro-inflammatory cytokine interleukin (IL)-8 from human airway epithelial cells through activation of PAR-2. Based on the knowledge that occupational airway symptoms are frequently presented by workers handling different species of fish and crustaceans [7,8,9,10,11,12,13,14,15], we have tested several types of seafood trypsins in our cell based assays. This in order to investigate possible initiation of signal transduction connected to inflammation processes in human airway epithelial cells [6,16]. During purification of numerous fish trypsins (Atlantic salmon [Salmo salar], sardine [Sardinops melanostictus], anchovy [Engraulis japonicus], jacopever [Sebastes schlegelii], yellow tail [Seriola quinqueradiata], spotted mackerel [Scomber australasicus] ) and trypsin from the king crab (Paralithodes camtschaticus) by fast protein liquid chromatography (FPLC), we observed that king crab trypsin bound stronger to the anionic column compared to the fish trypsins we purified
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