Digestive proteolysis in the Colorado potato beetle, Leptinotarsa decemlineata: Activity-based profiling and imaging of a multipeptidase network

Srp, Jaroslav; Nussbaumerová, Martina; Horn, Martin; Mareš, Michael

doi:10.1016/j.ibmb.2016.08.004

Cited by 12 publications

(15 citation statements)

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“…Recombinant SmSP2 expressed in P . pastoris (1 μg) was incubated 1 h at 37°C with 1 μM activity-based probe BoRC [ 35 ] in 50 mM Tris-HCl, pH 8.0. The competitive labeling reaction was also performed after treatment of SmSP2 for 15 min at 37°C with 1 mM serine protease inhibitor Pefabloc SC (Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

“…The active enzyme cleaved Z-F-R-AMC and migrated on SDS-PAGE as a single band of approximately 28 kDa ( Fig 3A ), consistent with the expected molecular mass of 30 kDa. rSmSP2 was visualized using the activity-based probe Bodipy-F-P-R-Cmk (BoRC) ( Fig 3A ) [ 35 ]; labeling by the probe was prevented by pre-incubation of the enzyme with the serine protease inhibitor, Pefabloc SC. Rabbit polyclonal antibodies, raised against the SmSP2 protease domain expressed in E .…”

Section: Homology Model Of Smsp2 Reveals the Trypsin-like Active Sitementioning

confidence: 99%

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SmSP2: A serine protease secreted by the blood fluke pathogen Schistosoma mansoni with anti-hemostatic properties

et al. 2018

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BackgroundSerine proteases are important virulence factors for many pathogens. Recently, we discovered a group of trypsin-like serine proteases with domain organization unique to flatworm parasites and containing a thrombospondin type 1 repeat (TSR-1). These proteases are recognized as antigens during host infection and may prove useful as anthelminthic vaccines, however their molecular characteristics are under-studied. Here, we characterize the structural and proteolytic attributes of serine protease 2 (SmSP2) from Schistosoma mansoni, one of the major species responsible for the tropical infectious disease, schistosomiasis.Methodology/Principal findingsSmSP2 comprises three domains: a histidine stretch, TSR-1 and a serine protease domain. The cleavage specificity of recombinant SmSP2 was determined using positional scanning and multiplex combinatorial libraries and the determinants of specificity were identified with 3D homology models, demonstrating a trypsin-like endopeptidase mode of action. SmSP2 displayed restricted proteolysis on protein substrates. It activated tissue plasminogen activator and plasminogen as key components of the fibrinolytic system, and released the vasoregulatory peptide, kinin, from kininogen. SmSP2 was detected in the surface tegument, esophageal glands and reproductive organs of the adult parasite by immunofluorescence microscopy, and in the excretory/secretory products by immunoblotting.Conclusions/SignificanceThe data suggest that SmSP2 is secreted, functions at the host-parasite interface and contributes to the survival of the parasite by manipulating host vasodilatation and fibrinolysis. SmSP2 may be, therefore, a potential target for anti-schistosomal therapy.

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Section: Methodsmentioning

confidence: 99%

Section: Homology Model Of Smsp2 Reveals the Trypsin-like Active Sitementioning

confidence: 99%

SmSP2: A serine protease secreted by the blood fluke pathogen Schistosoma mansoni with anti-hemostatic properties

et al. 2018

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“…Similar data are also available from model insects like Drosophila melanogaster (Dutta et al, 2015), Tribolium castaneum (Morris et al, 2009) and Manduca sexta (Pauchet et al, 2010). In comparison with medically important insects, agricultural Digestive proteases and pest responses to protease inhibitors or proteolytically activated bacterial toxins have received special attentions due to close associations with practical problems and biotechnological applications (Khajuria et al, 2010;Srp et al, 2016;Oppert et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Digestion-related proteins in the tobacco hornworm, Manduca sexta

Miao

Cao

Jiang

2020

Insect Biochemistry and Molecular Biology

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“…Cathepsin D evolved to an enzyme with digestive function in invertebrates like insects (Srp et al, 2016), mites (Wajahat Mahmood et al, 2013) and bloodsucking parasites as part of a multienzyme proteolytic complex (Sojka et al, 2016) and in vertebrates like bloodsucking fish (Xiao et al, 2015). How a typical intra-cellular lysosomal enzyme can appear in the gastric fluid of crustacea and act there as an extra-cellular enzyme (Rojo et al, 2010a) may be answered by the function and cytology of the digestive organs.…”

Section: Discussionmentioning

confidence: 99%

“…Cathepsin D evolved to an enzyme with digestive function in invertebrates like insects (Srp et al, 2016), mites (Wajahat Mahmood et al, 2013) and bloodsucking parasites as part of a multienzyme proteolytic complex (Sojka et al, 2016) and in vertebrates like bloodsucking fish (Xiao et al, 2015) and now we found it in crustaceans (Navarrete del Rojo et al, 2010a). The presence of cathepsin D in the gastric fluid of clawed lobsters that contributes to the extracellular digestion (Rojo et al, 2010b) raises the question whether this situation is unique within the genus of clawed lobsters (Homarus sp.)…”

Section: Introductionmentioning

confidence: 99%

Is digestive cathepsin D the rule in decapod crustaceans?

Martinez-Alarcon

Saborowski

Rojo‐Arreola

et al. 2018

Comparative Biochemistry and Physiology Part B: Biochemistry an

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A B S T R A C TCathepsin D is an aspartic endopetidase with typical characteristics of lysosomal enzymes. Cathepsin D activity has been reported in the gastric fluid of clawed lobsters where it acts as an extracellular digestive enzyme. Here we investigate whether cathepsin D is unique in clawed lobsters or, instead, common in decapod crustaceans. Eleven species of decapods belonging to six infraorders were tested for cathepsin D activity in the midgut gland, the muscle tissue, the gills, and when technically possible, in the gastric fluid. Cathepsin D activity was present in the midgut gland of all 11 species and in the gastric fluid from the seven species from which samples could be taken. All sampled species showed higher activities in the midgut glands than in non-digestive organs and the activity was highest in the clawed lobster. Cathepsin D mRNA was obtained from tissue samples of midgut gland, muscle, and gills. Analyses of deduced amino acid sequence confirmed molecular features of lysosomal cathepsin D and revealed high similarity between the enzymes from Astacidea and Caridea on one side, and the enzymes from Penaeoidea, Anomura, and Brachyura on the other side. Our results support the presence of cathepsin D activity in the midgut glands and in the gastric fluids of several decapod species suggesting an extracellular function of this lysosomal enzyme. We discuss whether cathepsin D may derive from the lysosomal-like vacuoles of the midgut gland B-cells and is released into the gastric lumen upon secretion by these cells.

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Digestive proteolysis in the Colorado potato beetle, Leptinotarsa decemlineata: Activity-based profiling and imaging of a multipeptidase network

Cited by 12 publications

References 46 publications

SmSP2: A serine protease secreted by the blood fluke pathogen Schistosoma mansoni with anti-hemostatic properties

SmSP2: A serine protease secreted by the blood fluke pathogen Schistosoma mansoni with anti-hemostatic properties

Digestion-related proteins in the tobacco hornworm, Manduca sexta

Is digestive cathepsin D the rule in decapod crustaceans?

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