Profiling of proteins and proteases in the products of the salivary gland, digestive tract and excretions from larvae of the camel nasal botfly, Cephalopina titillator (Clark)

2015 ◽  
Vol 70 (7-8) ◽  
pp. 197-203
Author(s):  
Hesham A. Yousef ◽  
Amira Afify ◽  
Afaf Abdel Meguid ◽  
Hany M. Hassan
Keyword(s):  
Salivary Gland ◽  
Digestive Tract ◽  
Serine Proteases ◽  
Protein Profiles ◽  
Zymogram Analysis ◽  
Larval Stages ◽  
Proteolytic Activities ◽  
Secretory Products

Abstract Proteins and proteolytic activities in the contents of the salivary gland (SGc), digestive tract (DTc) and excretory-secretory products (ESP) from larvae of the camel nasal botfly Cephalopina titillator were separated electrophoretically, and characterized. The protein profiles of the different samples were qualitatively quite similar in the larval stages L2 and L3. Zymogram analysis of proteases in the samples indicated that the digestive tract contained a greater variety of proteases than the salivary gland or the excretory-secretory products. They are mainly serine proteases. Proteases of ESP and DTc (especially of 3rd instar) contain trypsin- and chymotrypsin-like serine proteases, while the serine proteases of SGc are not of the trypsin- or chemotrypsin-type.

An initial characterization of the proteolytic enzymes secreted by the adult stage of the human hookworm Necator americanus

Parasitology ◽  
1995 ◽  
Vol 110 (5) ◽  
pp. 555-563 ◽  
Author(s):  
A. Brown ◽  
J. M. Burleigh ◽  
E. E. Billett ◽  
D. I. Pritchard
Keyword(s):  
Serine Proteases ◽  
Metal Ion ◽  
Proteolytic Enzymes ◽  
Maximum Activity ◽  
Biologically Relevant ◽  
Necator Americanus ◽  
Recent Developments ◽  
Proteolytic Activities ◽  
Secretory Products ◽  
Ph Range

SUMMARYThe proteolytic activities present in adult Necator americanus excretory–secretory products have been assessed using biologically relevant, naturally occurring substrates (haemoglobin and fibrinogen) and a number of synthetic fluorogenic and chromogenic substrates. One broad peak of activity was observed against haemoglobin in the pH range 5 to 7, with maximum activity at pH 6·6, while fibrinogenolytic activity was shown to be greater at pH 3·5. Inhibition studies against haemoglobin, fibrinogen and synthetic substrates using a battery of appropriate protease inhibitors indicated the presence of a mixture of aspartyl, cysteinyl and serine proteases. Metal ion (Ca2+, Zn2+ and Fe2+) stimulation was demonstrated, with stimulation by Zn2+ being the most marked. These results are discussed in the context of recent developments in the field of parasite proteolytic enzymes, where they have been suggested as targets for immuno- and chemotherapy.

scholarly journals Occurrence of Tetracampos ciliotheca and Proteocephalus glanduligerus in Clarias gariepinus (Burchell, 1822) collected from the Vaal Dam, South Africa

2013 ◽  
Vol 80 (1) ◽  
Author(s):  
Grace Madanire-Moyo ◽  
Annemariè Avenant-Oldewage
Keyword(s):  
South Africa ◽  
Digestive Tract ◽  
Standard Length ◽  
Feeding Habits ◽  
Clarias Gariepinus ◽  
Male And Female ◽  
Mean Intensity ◽  
Gill Nets ◽  
Larval Stages ◽  
Vaal Dam

Cestodes are parasitic flatworms that live in the digestive tract of vertebrates as adults and often in the liver, muscle, haemocoel, mesentery and brain of various animals as larval stages. To identify the cestodes infecting Clarias gariepinus Burchell, 1822 (sharptooth catfish) in the Vaal Dam, a total of 45 host specimens were collected with the aid of gill nets between October 2011, January and April 2012. The fish were sacrificed and examined for cestode parasites. Two adult cestodes, Tetracampos ciliotheca Wedl, 1861 (prevalence 86.7%, mean intensity = 15, n = 45) and Proteocephalus glanduligerus (Janicki, 1928) (prevalence 51.1%, mean intensity = 5, n = 45) were found in the intestines of the catfish. Both T. ciliotheca and P. glanduligerus are new locality records. There were statistically insignificant differences in the infection of the male and female C. gariepinu. Fish with standard length ranging from 40 cm – 54 cm (≥ 3 years) had the highest prevalence and mean intensity while those ranging from 10 cm – 24 cm (< 1 year) had the lowest prevalence and mean intensity for both cestodes. The study highlights the importance of changing feeding habits of C. gariepinus with age on the prevalence and mean intensity of the two gastrointestinal cestode parasites.

Study of the Proteolytic Activity of the Tropical Legume Crotalaria spectabilis

2012 ◽  
Vol 67 (9-10) ◽  
pp. 495-509 ◽  
Author(s):  
Juliana da Silva Pacheco ◽  
Raquel Elisa da Silva-Lopez
Keyword(s):  
Proteolytic Activity ◽  
Serine Proteases ◽  
Ph Value ◽  
Cysteine Proteases ◽  
Leaf Extracts ◽  
Serine Protease Activity ◽  
Proteolytic Activities ◽  
Low Levels ◽  
Crotalaria Spectabilis

The characterization of legume proteases contributes to the understanding of the physiology of plants and their interaction with the environment. Thirteen extracts from various parts of Crotalaria spectabilis were made using different extraction systems. The highest protein content was found in seeds, and the most pronounced proteolytic activity was observed in leaf extracts, with an optimal pH value in the alkaline range. Proteases in extracts from roots, stems, and fl owers were active in various pH ranges. Proteases in all extracts were maximally active between 30 °C and 60 °C and were thermostable (24 h, 60 °C). Hemoglobin, bovine serum albumin, casein, and gelatin were hydrolyzed by C. spectabilis extracts in different ways. The highest serine protease activity was found in leaves. Seeds contained high levels of serine proteases and low levels of cysteine proteases. Flowers, roots, and stems contained different levels of serine, aspartic, and metalloproteases, respectively. The proteolytic activities in extracts were modulated by cations and oxidants to various degrees. C. spectabilis proteases are differentially expressed in distinctive organs, and their stability against heat and oxidants makes this plant an important source of stable proteases

Protease activities of Acanthamoeba polyphaga and Acanthamoeba castellanii

2006 ◽  
Vol 52 (1) ◽  
pp. 16-23 ◽  
Author(s):  
José de Jesús Serrano-Luna ◽  
Isaac Cervantes-Sandoval ◽  
Jesús Calderón ◽  
Fernando Navarro-García ◽  
Victor Tsutsumi ◽  
...  
Keyword(s):  
Protease Activity ◽  
Serine Proteases ◽  
Biochemical Characterization ◽  
Acanthamoeba Castellanii ◽  
Cysteine Proteases ◽  
Skin Lesions ◽  
Sds Page ◽  
Serine Protease Family ◽  
Proteolytic Activities ◽  
Amoebic Keratitis

Acanthamoeba spp. are free-living amoebae that cause amoebic granulomatous encephalitis, skin lesions, and ocular amoebic keratitis in humans. Several authors have suggested that proteases could play a role in the pathogenesis of these diseases. In the present work, we performed a partial biochemical characterization of proteases in crude extracts of Acanthamoeba spp. and in conditioned medium using 7.5% SDS–PAGE copolymerized with 0.1% m/v gelatin as substrate. We distinguished a total of 17 bands with proteolytic activity distributed in two species of Acanthamoeba. The bands ranged from 30 to 188 kDa in A. castellanii and from 34 to 144 kDa in A. polyphaga. Additionally, we showed that the pattern of protease activity differed in the two species of Acanthamoeba when pH was altered. By using protease inhibitors, we found that the proteolytic activities belonged mostly to the serine protease family and secondly to cysteine proteases and that the proteolytic activities from A. castellanii were higher than those in A. polyphaga. Furthermore, aprotinin was found to in hibit crude extract protease activity on Madin–Darby canine kidney (MDCK) monolayers. These data suggest that protease patterns could be more complex than previously reported.Key words: Acanthamoeba spp., amoebic keratitis, serine proteases, cysteine proteases, cytopathic effect.

Some characteristics of extracellular proteases produced by members of the Chytridiales and the Spizellomycetales (Chytridiomycetes)

1994 ◽  
Vol 40 (2) ◽  
pp. 106-112 ◽  
Author(s):  
Thomas Krarup ◽  
Lauritz W. Olson ◽  
Hans Peter Heldt-Hansen
Keyword(s):  
Proteolytic Activity ◽  
Isoelectric Focusing ◽  
Serine Proteases ◽  
Proteolytic Enzymes ◽  
Complex Compounds ◽  
Extracellular Proteases ◽  
Peptide Substrates ◽  
Proteolytic Activities ◽  
Selection Of

The extracellular proteolytic enzymes of eight saprophytic, eucarpic, and monocentric isolates from two genera of the order Spizellomycetales and from one genus of the order Chytridiales (Chytridiomycetes) have been partially characterized. The isolectric points of the proteases were estimated from zymograms and demonstrate the existence of three types of proteolytic activity in most isolates. The proteases were tested against synthetic chromogenic peptide substrates and a selection of cations and more complex compounds, and the results suggest that parts of the extracellular proteolytic activities are due to proteases from two groups: the Ca2+ stabilized proteases and the alkaline serine proteases.Key words: serine proteases, metalloproteases, Chytridiomycetes, isoelectric focusing, chromogenic peptide substrates.

Formation of the gut in the first two naupliar stages of Acartia clausi and Hemidiaptomus roubaui (Copepoda, Calanoida): comparative structural and ultrastructural aspects

2002 ◽  
Vol 80 (2) ◽  
pp. 232-244 ◽  
Author(s):  
A Baud ◽  
R -M Barthélémy ◽  
S Nival ◽  
M Brunet
Keyword(s):  
Epithelial Cells ◽  
Digestive Tract ◽  
Digestive System ◽  
Larval Stages ◽  
Acartia Clausi ◽  
The Future ◽  
Endodermal Cells ◽  
Structure And Ultrastructure

In this study, the structure and ultrastructure of the digestive system are compared in the early larval stages (nauplii I and II) of two copepod calanoid species, Acartia clausi Giesbrecht, 1889 and Hemidiaptomus roubaui Richard, 1888. The nauplii I of both species have no functional digestive tract, which is represented initially only by a blind esophageal slit and yolky endodermal cells, which fill the most part of the naupliar body, whereas at the nauplius II stage the differentiated digestive tract becomes functional. The resorption cavity corresponding to the future midgut is progressively formed in the endodermal mass during the premolt phase; it is surrounded by differentiating epithelial cells. In the ecdysial phase the foregut has associated labral glands, the midgut young R-, B-, and R'-cells of epithelium, and there is a short open hindgut.

scholarly journals Positive selection on secretory and structural components of salivary glands within the ecologically diverse bat family Phyllostomidae

2019 ◽  
Author(s):  
Michael Vandewege ◽  
Cibele G. Sotero-Caio ◽  
Caleb D. Phillips
Keyword(s):  
Salivary Gland ◽  
Positive Selection ◽  
Salivary Glands ◽  
Single Gene ◽  
Go Annotation ◽  
Exact Function ◽  
Secretory Products ◽  
Cellular Components ◽  
Dietary Strategies ◽  
Extracellular Environment

Abstract The leaf-nosed bats (Phyllostomidae) are outliers among chiropterans with respect to the unusually high diversity of dietary strategies within the family. Salivary glands, owing to their functions and high ultrastructural variability among lineages, are proposed to have played an important role during the phyllostomid radiation. Salivary gland secretory products directly interact with food materials and pathogens which can provide selective interactions. To identify genes underlying salivary gland functional diversification, we sequenced submandibular gland transcriptomes from phyllostomid species representative of divergent dietary strategies. From the assembled transcriptomes, we identified and tested 3,266 single gene orthologs and identified 57 evolving under positive selection. All enriched gene ontology terms were related to defense against other organisms and the cell membrane/extracellular environment. Based on GO annotation, many of the defense-related loci under selection form secretory products. Salivary glands perform a complex array of tasks, and we identified positive selection on additional proteins bound to membranes and in the extracellular environment with multiple functions. Although illuminating exact function is difficult, results suggest that regulatory release of secretory products, not the products themselves, is disproportionately shaped by positive selection on coding sequence. Identified instances of selection on such cellular components may help explain the ultrastructural variability of salivary glands previously documented.

scholarly journals Proteinases in Excretory-Secretory Products ofToxocara canisSecond-Stage Larvae: Zymography and Modeling Insights

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Gonzalo Ernesto González-Páez ◽  
Fernando Alba-Hurtado ◽  
Carlos Gerardo García-Tovar ◽  
Raúl Argüello-García
Keyword(s):  
Serine Proteinase ◽  
Toxocara Canis ◽  
Serine Proteinases ◽  
Bodily Fluids ◽  
Multiple Components ◽  
Proteolytic Activities ◽  
Secretory Products ◽  
Ph Range ◽  
Physiological Substrates

Components released in excretory-secretory products ofToxocara canislarvae (TES) include phosphatidylethanolamine-binding proteins (TES26), mucins (TES120, MUC2-5), and C-type lectins (TES32, TES70) and their biochemical, immunological, and diagnostic properties have been extensively studied albeit proteinase activities towards physiological substrates are almost unknown. Proteolytic activities in TES samples were first analyzed by gel electrophoresis with gelatin as substrate. Major activities of ~400, 120, and 32 kDa in TES were relatively similar over a broad pH range (5.5–9.0) and all these were of the serine-type as leupeptin abolished gelatinolysis. Further, the ~400 kDa component degraded all physiological substrates tested (laminin, fibronectin, albumin, and goat IgG) and the 120 kDa component degraded albumin and goat IgG while proteinases of lower MW (45, 32, and 26 kDa) only degraded laminin and fibronectin, preferentially at alkaline pH (9.0). By protein modeling approaches using the known sequences of TES components, only TES26 and MUC4 displayed folding patterns significantly related to reference serine proteinases. These data suggest that most of serine proteinase activities secretedin vitroby infective larvae ofT. canishave intriguing nature but otherwise help the parasite to affect multiple components of somatic organs and bodily fluids within the infected host.

Structural and physicochemical changes due to proteolytic deterioration of escamoles (Liometopum apiculatum) a traditional Mexican food

2015 ◽  
Vol 1 (4) ◽  
pp. 271-280 ◽  
Author(s):  
A. Castillo-Andrade ◽  
R. García-Barrientos ◽  
M.A. Ruiz-Cabrera ◽  
C. Rivera-Bautista ◽  
J.D. Pérez-Martínez ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Serine Proteases ◽  
Structural Changes ◽  
Iodoacetic Acid ◽  
Initial Structure ◽  
Physicochemical Changes ◽  
Proteolytic Activities ◽  
Food Needs ◽  
Endogenous Proteases

Entomophagy or consumption of insects has significantly increased worldwide, either for pleasure or to satisfy the food needs in developing countries. There are approximately 2,000 species of edible insects distributed in 120 countries. From these 2,000 species, about 540 are located in Mexico; one of the most consumed are the escamoles. Escamoles are larvae and pupae of the ant Liometopum apiculatum. Escamoles are nutritious because of their high content of protein, fat, carbohydrates and vitamin. However, during storage the quality of escamoles changes rapidly which affects the acceptability by the consumer. This loss of quality is probably a result of proteolytic activity of endogenous proteases. Therefore, the objectives of this study were to identify the classes of proteases in escamoles as well as to evaluate the effect of proteolytic activity on physicochemical and structural changes during storage. Proteases identification was conducted using specific inhibitors; structural changes, texture, and proteolytic activity were monitored at different days of storage. The highest proteolytic activities (P<0.05) were observed at pH 8, 9 and 10 and at 37 and 50 °C. Proteases were mainly inhibited by iodoacetic acid and soybean trypsin inhibitor showing that cysteine and serine proteases were dominant. High proteolytic activity, significant (P<0.05) reduction in texture and weight loss was observed during storage. The deterioration of escamoles was evident in analyses of images, where initial structure was lost during storage. These results indicate that different groups of proteases are associated with rapid deterioration of escamoles.

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