scholarly journals Venomics and Cellular Toxicity of Thai Pit Vipers (Trimeresurus macrops and T. hageni)

Toxins ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 54 ◽  
Author(s):  
Supeecha Kumkate ◽  
Lawan Chanhome ◽  
Tipparat Thiangtrongjit ◽  
Jureeporn Noiphrom ◽  
Panithi Laoungboa ◽  
...  
Keyword(s):  
Snake Venom ◽  
Protein Composition ◽  
Treatment Method ◽  
Cross Reactivity ◽  
Major Protein ◽  
Dependent Manner ◽  
Crude Venom ◽  
Monocytic Cells ◽  
Protein Components ◽  
Species Specific

The two venomous pit vipers, Trimeresurus macrops and T. hageni, are distributed throughout Thailand, although their abundance varies among different areas. No species-specific antivenom is available for their bite victims, and the only recorded treatment method is a horse antivenom raised against T. albolabris crude venom. To facilitate assessment of the cross-reactivity of heterologous antivenoms, protein profiles of T. macrops and T. hageni venoms were explored using mass-spectrometry-based proteomics. The results show that 185 and 216 proteins were identified from T. macrops and T. hageni venoms, respectively. Two major protein components in T. macrops and T. hageni venoms were snake venom serine protease and metalloproteinase. The toxicity of the venoms on human monocytes and skin fibroblasts was analyzed, and both showed a greater cytotoxic effect on fibroblasts than monocytic cells, with toxicity occurring in a dose-dependent rather than a time-dependent manner. Exploring the protein composition of snake venom leads to a better understanding of the envenoming of prey. Moreover, knowledge of pit viper venomics facilitates the selection of the optimum heterologous antivenoms for treating bite victims.

Stage-specific patterns of collagen gene expression during development of Caenorhabditis elegans

1985 ◽  
Vol 5 (2) ◽  
pp. 363-372
Author(s):  
G N Cox ◽  
D Hirsh
Keyword(s):  
Caenorhabditis Elegans ◽  
Protein Composition ◽  
Large Family ◽  
Major Protein ◽  
Mrna Levels ◽  
Mrna Accumulation ◽  
C Elegans ◽  
Collagen Protein ◽  
Protein Components ◽  
Collagen Genes

Collagens are the major protein components of the Caenorhabditis elegans cuticle and are encoded by a large family of 40 to 150 closely related but nonidentical genes. We have determined temporal patterns of mRNA accumulation for a large number of collagen genes by screening recombinant phages and plasmids containing cloned collagen genes under high stringency conditions with 32P-labeled cDNA preparations specific for eggs or three postembryonic molts. We find that collagen mRNA levels are regulated both temporally and quantitatively during C. elegans development. Most genes studied exhibit one of four patterns of mRNA accumulation which correlate with changes in cuticle morphology and collagen protein composition during development. Our results suggest that, in general, there is a progressive activation of new collagen genes during normal development.

Protein composition and protein synthesis in the surface membranes of Schistosoma mansoni

Parasitology ◽  
1972 ◽  
Vol 65 (1) ◽  
pp. 55-69 ◽  
Author(s):  
J. R. Kusel
Keyword(s):  
Protein Synthesis ◽  
Calcium Chloride ◽  
Unit Area ◽  
Surface Membrane ◽  
Biological Significance ◽  
Protein Composition ◽  
Major Protein ◽  
Freezing And Thawing ◽  
Protein Components ◽  
Protein Incorporation

Saponin treatment of cercariae and schistosomula alters the surface membrane so that it may be sheared from the organism and isolated as fragments by centrifugation. Saponin-calcium chloride treatment or freezing and thawing of adult worms removes the surface membrane, which can be washed from the bodies and collected by centrifugation. The small quantities of material available necessitated the development of a sensitive technique for detecting the proteins in the membranes. The surface membranes were radioiodinated after butan-1-ol extraction and electrophoresed in polyacrylamide gel. Adult surfaces prepared by the saponin-calcium chloride and by the freezing and thawing technique had identical protein components, detected as gel-cut profiles or in autoradiographs. The quantity of a rapidly migrating PAS-positive amido black negative component was greater in the surfaces prepared by saponincalcium chloride than in the frozen and thawed surfaces. This component contains lipid, some of which may be glycolipid. It was largely absent from the surface membranes of cercariae and schistosomula. Cercarial surface membranes contained a major protein component which was absent from the surface membranes of schistosomula. Otherwise the surface membranes of the cercariae were identical to those of the schistosomula in their protein components. The rate of incorporation of freshly synthesized protein per unit area of surface membrane of schistosomula was very low in the first 5 days in culture, after which there was a very rapid increase to a maximum rate on the 15th day. After this time, the rate of protein synthesis decreased to a low level at 26 days. In these studies the activity per unit area was measured and this would not be expected to vary greatly during growth. The biological significance of the observed variation in protein incorporation into the membrane is unclear.

scholarly journals New Insights on Moojase, a Thrombin-Like Serine Protease from Bothrops moojeni Snake Venom

Toxins ◽  
2018 ◽  
Vol 10 (12) ◽  
pp. 500 ◽  
Author(s):  
Fernanda Amorim ◽  
Danilo Menaldo ◽  
Sante Carone ◽  
Thiago Silva ◽  
Marco Sartim ◽  
...  
Keyword(s):  
Snake Venom ◽  
Serine Proteases ◽  
New Drugs ◽  
Coagulation Cascade ◽  
Dependent Manner ◽  
Crude Venom ◽  
Human Fibrinogen ◽  
Factor Xia ◽  
Bothrops Moojeni

Snake venom serine proteases (SVSPs) are enzymes that are capable of interfering in various parts of the blood coagulation cascade, which makes them interesting candidates for the development of new therapeutic drugs. Herein, we isolated and characterized Moojase, a potent coagulant enzyme from Bothrops moojeni snake venom. The toxin was isolated from the crude venom using a two-step chromatographic procedure. Moojase is a glycoprotein with N-linked glycans, molecular mass of 30.3 kDa and acidic character (pI 5.80–6.88). Sequencing of Moojase indicated that it is an isoform of Batroxobin. Moojase was able to clot platelet-poor plasma and fibrinogen solutions in a dose-dependent manner, indicating thrombin-like properties. Moojase also rapidly induced the proteolysis of the Aα chains of human fibrinogen, followed by the degradation of the Bβ chains after extended periods of incubation, and these effects were inhibited by PMSF, SDS and DTT, but not by benzamidine or EDTA. RP-HPLC analysis of its fibrinogenolysis confirmed the main generation of fibrinopeptide A. Moojase also induced the fibrinolysis of fibrin clots formed in vitro, and the aggregation of washed platelets, as well as significant amidolytic activity on substrates for thrombin, plasma kallikrein, factor Xia, and factor XIIa. Furthermore, thermofluor analyses and the esterase activity of Moojase demonstrated its very high stability at different pH buffers and temperatures. Thus, studies such as this for Moojase should increase knowledge on SVSPs, allowing their bioprospection as valuable prototypes in the development of new drugs, or as biotechnological tools.

scholarly journals Composition and Acute Inflammatory Response from Tetraponera rufonigra Venom on RAW 264.7 Macrophage Cells

Toxins ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 257
Author(s):  
Suwatjanee Naephrai ◽  
Supakit Khacha-ananda ◽  
Pornsiri Pitchakarn ◽  
Churdsak Jaikang
Keyword(s):  
Protein Composition ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
Crude Venom ◽  
Venom Protein ◽  
Macrophage Cells ◽  
Raw 264.7 Macrophage ◽  
Ant Venom ◽  
Raw 264.7 Macrophage Cells ◽  
Cox 2

Tetraponera rufonigra (Arboreal Bicoloured Ant) venom induces pain, inflammation, and anaphylaxis in people and has an increased incident in Southeast Asia regions. The bioactive components and mechanism of action of the ant venom are still limited. The aim of this research was to identify the protein composition and inflammatory process of the ant venom by using RAW 264.7 macrophage cells. The major venom proteins are composed of 5’ nucleotidase, prolyl endopeptidase-like, aminopeptidase N, trypsin-3, venom protein, and phospholipase A2 (PLA2). The venom showed PLA2 activity and represented 0.46 μg of PLA2 bee venom equivalent/μg crude venom protein. The venom induced cytotoxic in a dose- and time-dependent manner with IC20 approximately at 4.01 µg/mL. The increased levels of COX-2 and PGE2 were observed after 1 h of treatment correlating with an upregulation of COX-2 expression. Moreover, the level of mPGES-1 expression was obviously increased after 12 h of venom induction. Hence, our results suggested that the induction of COX-2/mPGEs-1 pathway could be a direct pathway for the ant venom-induced inflammation.

scholarly journals Stage-specific patterns of collagen gene expression during development of Caenorhabditis elegans.

1985 ◽  
Vol 5 (2) ◽  
pp. 363-372 ◽  
Author(s):  
G N Cox ◽  
D Hirsh
Keyword(s):  
Caenorhabditis Elegans ◽  
Protein Composition ◽  
Large Family ◽  
Major Protein ◽  
Mrna Levels ◽  
Mrna Accumulation ◽  
C Elegans ◽  
Collagen Protein ◽  
Protein Components ◽  
Collagen Genes

Collagens are the major protein components of the Caenorhabditis elegans cuticle and are encoded by a large family of 40 to 150 closely related but nonidentical genes. We have determined temporal patterns of mRNA accumulation for a large number of collagen genes by screening recombinant phages and plasmids containing cloned collagen genes under high stringency conditions with 32P-labeled cDNA preparations specific for eggs or three postembryonic molts. We find that collagen mRNA levels are regulated both temporally and quantitatively during C. elegans development. Most genes studied exhibit one of four patterns of mRNA accumulation which correlate with changes in cuticle morphology and collagen protein composition during development. Our results suggest that, in general, there is a progressive activation of new collagen genes during normal development.

scholarly journals Neutrophil stimulation with citrullinated histone H4 slows down calcium influx and reduces NET formation compared with native histone H4

PLoS ONE ◽  
2021 ◽  
Vol 16 (5) ◽  
pp. e0251726
Author(s):  
Lai Shi ◽  
Karen Aymonnier ◽  
Denisa D. Wagner
Keyword(s):  
Extracellular Space ◽  
Ischemia Reperfusion Injury ◽  
Calcium Influx ◽  
Ischemia Reperfusion ◽  
Neutrophil Extracellular Trap ◽  
Major Protein ◽  
Dependent Manner ◽  
Histone H4 ◽  
Native Form ◽  
Protein Components

Peptidylarginine deiminase 4 (PAD4) catalyzes posttranslational modification of many target proteins through converting protein arginine or mono-methylarginine to citrulline. Neutrophil extracellular trap (NET) formation is the most dramatic manifestation of PAD4-mediated hypercitrullination reaction in neutrophils, which is characterized by the release of nuclear chromatin to form a chromatin network in the extracellular space. Histones H4, one of the major protein components of chromatin, is released into the extracellular space during sepsis, trauma, and ischemia-reperfusion injury and can also be released during the process of NET formation, along with its citrullinated form. The present study showed that histone H4 can induce NET formation in a calcium and PAD4 dependent manner. Histone H4 caused permeabilization of the neutrophil membrane and sustained rise in intracellular calcium that is necessary for activation of PAD4. In comparison, citrullinated histone H4 induced less calcium influx compared with its native form, leading to reduced NET formation. These studies suggest that citrullinated histone H4 could serve as a brake in the pathology of NETs, slowing down the vicious circle between histone H4 and NETs.

scholarly journals Identification and hemolytic activity of jellyfish (Rhopilema sp., Scyphozoa: Rhizostomeae) venom from the Persian Gulf and Oman Sea

2019 ◽  
Vol 20 (4) ◽  
pp. 1228-1232
Author(s):  
HOSSEIN JAFARI ◽  
HOSSEIN HONARI ◽  
JAMIL ZARGAN ◽  
SAEID TAMADONI JAHROMI
Keyword(s):  
Hemolytic Activity ◽  
Persian Gulf ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Biologically Active ◽  
Major Protein ◽  
Crude Venom ◽  
Oman Sea ◽  
Protein Components ◽  
The Persian Gulf

Abstract. Jafari H, Honari H, Zargan J, Jahromi ST. 2019. Identification and hemolytic activity of jellyfish (Rhopilema sp., Scyphozoa: Rhizostomeae) venom from the Persian Gulf and Oman Sea. Biodiversitas 20: 1228-1232. The present study investigated the hemolytic capacity of the crude venom extracted from isolated nematocysts of Rhopilema sp. Scyphozoa: Rhizostomeae. Nematocyst was used at various concentrations to evaluate the hemolytic activity by using the nematocysts of human, mice, and sheep. Mean concentration-dependent hemolysis could be observed from 200 µg/mL of protein equivalents or higher with variable potencies in different species. The crude venom was analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis SDS-PAGE. Molecular weight with 3 clear bands including 45, 65 and 95 kDa appeared to be the major protein components of the venom. The results of our experiments indicated that venom of Rhopilema sp. induces hemolysis in the studied species and determined that the increase in the amount of toxin has a positive correlation with the increase of cell lysis. This study showed that the venom of Rhopilema sp. may have many biologically active principles, which need further studies in the future.

Trimucytin: A Collagen-Like Aggregating Inducer Isolated from Trimeresurus mucrosquamatus Snake Venom

1993 ◽  
Vol 69 (03) ◽  
pp. 286-292 ◽  
Author(s):  
Che-Ming Teng ◽  
Feng-Nien Ko ◽  
Inn-Ho Tsai ◽  
Man-Ling Hung ◽  
Tur-Fu Huang
Keyword(s):  
Platelet Aggregation ◽  
Snake Venom ◽  
Inositol Phosphate ◽  
Shape Change ◽  
Platelet Activating Factor ◽  
Atp Release ◽  
Platelet Membrane ◽  
Thromboxane B2 ◽  
Dependent Manner ◽  
Concentration Dependent Manner

SummaryTrimucytin is a potent platelet aggregation inducer isolated from Trimeresurus mucrosquamatus snake venom. Similar to collagen, trimucytin has a run of (Gly-Pro-X) repeats at the N-terminal amino acids sequence. It induced platelet aggregation, ATP release and thromboxane formation in rabbit platelets in a concentration-dependent manner. The aggregation was not due to released ADP since it was not suppressed by creatine phosphate/creatine phosphokinase. It was not either due to thromboxane A2 formation because indomethacin and BW755C did not have any effect on the aggregation even thromboxane B2 formation was completely abolished by indomethacin. Platelet-activating factor (PAF) was not involved in the aggregation since a PAF antagonist, kadsurenone, did not affect. However, RGD-containing peptide triflavin inhibited the aggregation, but not the release of ATP, of platelets induced by trimucytin. Indomethacin, mepacrine, prostaglandin E1 and tetracaine inhibited the thromboxane B2 formation of platelets caused by collagen and trimucytin. Forskolin and sodium nitroprusside inhibited both platelet aggregation and ATP release, but not the shape change induced by trimucytin. In quin-2 loaded platelets, the rise of intracellular calcium concentration caused by trimucytin was decreased by 12-O-tetradecanoyl phorbol-13 acetate, imipramine, TMB-8 and indomethacin. In the absence of extracellular calcium, both collagen and trimucytin caused no thromboxane B2 formation, but still induced ATP release which was completely blocked by R 59022. Inositol phosphate formation in platelets was markedly enhanced by trimucytin and collagen. MAB1988, an antibody against platelet membrane glycoprotein Ia, inhibited trimucytinand collagen-induced platelet aggregation and ATP release. However, trimucytin did not replace the binding of 125I-labeled MAB1988 to platelets. Platelets pre-exposed to trimucytin were resistant to the second challenge with trimucytin itself or collagen. It is concluded that trimucytin may activate collagen receptors on platelet membrane, and cause aggregation and release mainly through phospholipase C-phosphoinositide pathway.

Impairment of Platelet Aggregation by Echis colorata Venom Mediated by L-Amino Acid Oxidase or H2O2

1982 ◽  
Vol 48 (03) ◽  
pp. 277-282 ◽  
Author(s):  
I Nathan ◽  
A Dvilansky ◽  
T Yirmiyahu ◽  
M Aharon ◽  
A Livne
Keyword(s):  
Hydrogen Peroxide ◽  
Amino Acid ◽  
Platelet Aggregation ◽  
Snake Venom ◽  
Oxidase Activity ◽  
Enzyme Preparation ◽  
Acid Oxidase ◽  
Crude Venom ◽  
Amino Acid Oxidase ◽  
Hemostatic Disorders

SummaryEchis colorata bites cause impairment of platelet aggregation and hemostatic disorders. The mechanism by which the snake venom inhibits platelet aggregation was studied. Upon fractionation, aggregation impairment activity and L-amino acid oxidase activity were similarly separated from the crude venom, unlike other venom enzymes. Preparations of L-amino acid oxidase from E.colorata and from Crotalus adamanteus replaced effectively the crude E.colorata venom in impairment of platelet aggregation. Furthermore, different treatments known to inhibit L-amino acid oxidase reduced in parallel the oxidase activity and the impairment potency of both the venom and the enzyme preparation. H2O2 mimicked characteristically the impairment effects of L-amino acid oxidase and the venom. Catalase completely abolished the impairment effects of the enzyme and the venom. It is concluded that hydrogen peroxide formed by the venom L-amino acid oxidase plays a role in affecting platelet aggregation and thus could contribute to the extended bleeding typical to persons bitten by E.colorata.

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