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Yoo Jung Park Ha Young Lee Young Su Jung Joon Seong Park Jae Sam Hwang Yoe-Sik Bae 《BMB reports》2015,48(8):479-484
In this study, we report that one of the antimicrobial peptides scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates actin polymerization and the subsequent chemotactic migration of macrophages through the activation of ERK and protein kinase B (Akt) activity. The scolopendrasin VII-induced chemotactic migration of macrophages is inhibited by the formyl peptide receptor 1 (FPR1) antagonist cyclosporine H. We also found that scolopendrasin VII stimulate the chemotactic migration of FPR1-transfected RBL-2H3 cells, but not that of vector-transfected cells; moreover, scolopendrasin VII directly binds to FPR1. Our findings therefore suggest that the antimicrobial peptide scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates macrophages, resulting in chemotactic migration via FPR1 signaling, and the peptide can be useful in the study of FPR1-related biological responses. [BMB Reports 2015; 48(8): 479-484] 相似文献
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Purification and molecular cloning of a novel serine protease from the centipede, Scolopendra subspinipes mutilans 总被引:5,自引:0,他引:5
You WK Sohn YD Kim KY Park DH Jang Y Chung KH 《Insect biochemistry and molecular biology》2004,34(3):239-250
A novel serine protease, named as scolonase, was purified and characterized from the tissue of the Korean centipede, Scolopendra subspinipes mutilans. Purified scolonase showed an apparent molecular weight of 25 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and an isoelectric point of 4.8 on isoelectric focusing gel. Scolonase was able to preferentially hydrolyze arginine over lysine at the cleavage site among the several synthetic peptide substrates. Scolonase has also a potent fibrinolytic activity by converting human Glu-plasminogen to activated plasmin due to the specific cleavage of the molecule at the peptide bond Arg(561)-Val(562). The enzyme activity of scolonase was completely inhibited by phenylmethanesulfonyl fluoride and difluorophosphate. The cDNA encoding scolonase was cloned from the cDNA library of the centipede constructed with oligonucleotide probe, which was designed on the basis of the N-terminal amino acid sequence of scolonase. The deduced complete amino acid sequence of scolonase demonstrated that the protein is composed of 277 amino acids including 33 amino acids as a leader sequence, and that it has significant sequence homology with other serine proteases. 相似文献
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Many animal phyla contain clades in which most or all species are venom-injecting predators. An example, in the arthropods, is the class Chilopoda, containing the approximately 3500 species of centipedes. Very little ecological or behavioural work yielding quantitative data has been conducted on centipede predation. Here, we describe a study of this kind. Our experiments employed one centipede species - a large tropical one, Scolopendra subspinipes mutilans - and two species of prey - a cricket, Gryllus assimilis, and a locust, Schistocerca gregaria. We conducted two experiments. The first was aimed at investigating the extent to which the centipedes attacked prey in particular tagmata as opposed to at random over the whole body surface. The results showed that the centipedes were highly selective, preferring to attack the head or thorax rather than the abdomen; indeed, they often reoriented the prey in order to achieve this. A possible explanation of this behaviour is to maximize the speed with which the neurotoxins in the venom reach either the brain or the thoracic ganglia that control limb movement. The second experiment was aimed at investigating the effect of venom-extraction on the attack rate, and specifically at testing if the magnitude of any such effect differed between the two types of prey, which differ considerably in size. The results showed a major effect of venom extraction in relation to both types of prey, but with the time taken to return to a 'normal' attack rate being longer in the case of the larger prey-type, namely the locust. We discuss these results in relation to the 'venom optimization hypothesis' and, more generally, to the principle of minimizing the production/use of venom, which is an energetically expensive resource. 相似文献
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Xuelai He Kwang Sik Lee Bo Yeon Kim Kyeong Yong Lee Hyeon Jin Ko Jingming Jia Hyung Joo Yoon Byung Rae Jin 《Journal of Asia》2019,22(1):19-24
Centipede venom contains a variety of proteins, peptides, and enzymes. However, the biological actions of toxin peptides in centipede venom remain largely unknown. In this study, we identified a centipede (Scolopendra subspinipes mutilans) venom toxin peptide (SsmTP) that was shown to act as a cell growth factor at low concentrations-in vitro. SsmTP was found to consist of 66 amino acids that display seven cysteine residues, which exhibited high similarity to the predicted neurotoxin. SsmTP was expressed in the venom gland of S. s. mutilans. A recombinant SsmTP peptide of approximately 5.2 kDa was produced in baculovirus-infected insect cells. Interestingly, SsmTP exhibited cytotoxicity in murine cells in a concentration-dependent manner but displayed a cell growth effect at low concentrations. SsmTP at a low concentration also protected murine cells against oxidative damage through the inhibition of caspase-1 and apoptosis. Our data indicate that SsmTP acts as a cell growth factor and a toxin peptide in a concentration-dependent manner. Consequently, our results provide evidence that the identification of SsmTP, a centipede toxin peptide, may not only elucidate the biological action of toxin peptides but also offer additional insight into the pharmacological applications of centipede venoms. 相似文献
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少棘巨蜈蚣(ScolopendrasubspinipesmutilansL.Koch)经95%乙醇脱脂后,再经4℃水冷渗,水提液低温旋转浓缩,冻干,得到的冻干粉先后经过SephadexG-25柱,等电聚焦制备电泳,再经SephadexG-150柱,SephadexG-100柱,最后经HPLC制备得到一个纯的碱性蛋白,命名为SSmp-d.该蛋白经HPLC、超薄等电聚焦电泳检验是均一的.采用HPLC和Protein-PakTM125柱测定其分子量为24.64kD.IEF-HPCE显示其等电点为9.27.氨基酸分析表明SSmp-d含较多的Arg、Lys等碱性氨基酸,另外还含有较多的Ala、Leu.使用蛋白质自动序列分析仪测定了SSmp-dN端的11个氨基酸,序列为NH3+-Asp-Val-Asn-Phe-Arg-Leu-Ser-Gly-Ala-Asp-Pro. 相似文献
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Many studies have been performed on venomous peptides derived from animals. However, little of this research has focused on peptides from centipede venoms. Here, a venom gland cDNA library was suc-cessfully constructed for the centipede Scolopendra subspinipes mutilans. A new cDNA encoding the precursor of a venom peptide, named SsmTx, was cloned from the venomous gland cDNA library of the centipede S. subspinipes mutilans. The full-length SsmTx cDNA sequence is 465 nt, including a 249 nt ORF, a 45 nt 5′ UTR and a 171 nt 3′ UTR. There is a signal tail AATAAA 31 nt upstream of the poly (A) tail. The precursor nucleotide sequence of SsmTx encodes a signal peptide of 25 residues and a mature peptide of 57 residues, which is bridged by two pairs of disulfide bonds. SsmTx displays a unique cysteine motif that is completely different from that of other venomous animal toxins. This is the first reported cDNA sequence encoding a venom peptide from the centipede S. subspinipes mutilans. 相似文献
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蜈蚣(Scolopendra subspinipes mutilans L.Koch)毒的化学组成和生物活性 总被引:1,自引:0,他引:1
系统分析少棘蜈蚣粗毒的化学组成,蛋白质含量86.23%、水不溶物质0.24%、还原糖0.23%、水份2.1%,仅含Ser、Pro、Arg等三种游离氨基酸,鉴定了11种微量元素,碱性凝胶电泳显示20条蛋白质谱带。分析了10种酶的活性、出血毒性及对血小板聚集的影响,其中精氨酸酯酶活力最高,不存在类凝血酶、淀粉酶活性及出血毒性,蜈蚣毒的浓度为0.3μg/μL时强烈诱导血小板的聚集。 相似文献
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Michel M. Dugon Luke Hayden Alexander Black Wallace Arthur 《Evolution & development》2012,14(6):515-521
In contrast to previous claims that (a) there is a law of recapitulation and, conversely, (b) recapitulation never happens, the evolutionary repatterning of development can take many forms, of which recapitulation is one. Here, we add another example to the list of case studies of recapitulation. This example involves the development of the venom claws (forcipules) in the centipede Scolopendra subspinipes mutilans, and in particular the development of the duct through which venom flows from the gland that produces it (proximal) to the opening called the meatus (distal) through which it is injected into prey. Most of the information we present is from early postembryonic stages—these have been neglected in previous work on centipede development. We show that the venom ducts arise from sutures that are invaginations of the cuticle. In S. s. mutilans, the invagination in each forcipule forms into a tubular structure that detaches itself from the exoskeleton and moves toward the center of the forcipule. This is in contrast to extant Scutigera, and also, probably, Scolopendra's extinct Scutigera‐like ancestors, where the duct remains attached to the cuticle of throughout development. Thus, S. s. mutilans exhibits a recapitulatory repatterning of development. 相似文献
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Scolopendra subspinipes mutilans, also known as Chinese red‐headed centipede, is a venomous centipede from East Asia and Australasia. Venom from this animal has not been researched as thoroughly as venom from snakes, snails, scorpions, and spiders. In this study, we isolated and characterized SsmTx‐I, a novel neurotoxin from the venom of S. subspinipes mutilans. SsmTx‐I contains 36 residues with four cysteines forming two disulfide bonds. It had low sequence similarity (<10%) with other identified peptide toxins. By whole‐cell recording, SsmTx‐I significantly blocked voltage‐gated K+ channels in dorsal root ganglion neurons with an IC50 value of 200 nM, but it had no effect on voltage‐gated Na+ channels. Among the nine K+ channel subtypes expressed in human embryonic kidney 293 cells, SsmTx‐I selectively blocked the Kv2.1 current with an IC50 value of 41.7 nM, but it had little effect on currents mediated by other K+ channel subtypes. Blockage of Kv2.1 by SsmTx‐I was not associated with significant alteration of steady‐state activation, suggesting that SsmTx‐I might act as a simple inhibitor or channel blocker rather than a gating modifier. Our study reported a specific Kv2.1‐blocker from centipede venom and provided a basis for future investigations of SsmTx‐I, for example on structure–function relationships, mechanism of action, and pharmacological potential. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
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The centipede Scolopendra subspinipes mutilans has been a medically important arthropod species by using it as a traditional medicine for the treatment of various diseases. In this study, we derived a novel lactoferricin B like peptide (LBLP) from the whole bodies of adult centipedes, S. s. mutilans, and investigated the antifungal effect of LBLP. LBLP exerted an antifungal and fungicidal activity without hemolysis. To investigate the antifungal mechanism of LBLP, a membrane study with propidium iodide was first conducted against Candida albicans. The result showed that LBLP caused fungal membrane permeabilization. The assays of the three dimensional flow cytometric contour plot and membrane potential further showed cell shrinkage and membrane depolarization by the membrane damage. Finally, we confirmed the membrane-active mechanism of LBLP by synthesizing model membranes, calcein and FITC-dextran loaded large unilamellar vesicles. These results showed that the antifungal effect of LBLP on membrane was due to the formation of pores with radii between 0.74 nm and 1.4 nm. In conclusion, this study suggests that LBLP exerts a potent antifungal activity by pore formation in the membrane, eventually leading to fungal cell death. 相似文献
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Expression,purification and functional characterization of a recombinant scorpion venom peptide BmTXKbeta 总被引:1,自引:0,他引:1
BmTXKbeta, a scorpion toxin isolated from the Chinese scorpion Buthus martensii Karsch (BmK), was expressed as a GST fusion protein in BL21 (DE3) strain. The recombinant GST-BmTXKbeta protein was purified by affinity chromatography. When treated with enterokinase, the GST-BmTXKbeta fusion protein released an approximate 6.5kDa protein which was the expected size for correctly processed. About 2mg purified recombinant BmTXKbeta protein (rBmTXKbeta) was produced from 1l bacterial culture, using this expression and purification system. The function of rBmTXKbeta was studied on the rabbit atrial myocyte by whole-cell patch clamp technique. The results showed that rBmTXKbeta inhibited the transient outward current (I(to)) of rabbit atrial myocyte with recovery after washout and the inhibition was concentration-dependent. The rBmTXKbeta prolonged the action potential duration of rabbit atrial myocyte in a concentration-dependent manner, whereas it did not affect the action potential amplitude. 相似文献
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J. G. E. Lewis 《Journal of Zoology》1966,149(2):188-203
The life history, reproduction, food and parasites of a population of Scolopendra amazonica Bücherl from the south bank of the Blue Nile at Khartoum, Sudan are described and discussed and Sudanese specimens are compared with Indian S. amazonica and Scolopendra morsitans. 相似文献
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Xu X Jin F Yu X Ji S Wang J Cheng H Wang C Zhang W 《Protein expression and purification》2007,53(2):293-301
Insect cecropins are small basic polypeptides synthesized in fat body and hemocytes in response to bacterial infections or hypodermic injuries. To explore a new approach for high expression of soluble cecropin in Escherichia coli cells, we fused the sequence encoding Musca domestica mature cecropin (named Mdmcec) in-frame to thioredoxin (TRX) gene to construct an expression vector pTRX-6His-Mdmcec. An enterokinase cleavage site was introduced between the 6xHis-tag and Mdmcec to facilitate final release of the recombinant Mdmcec. The fusion protein TRX-6His-Mdmcec was purified successfully by HisTrap HP affinity column and a high yield of 48.0mg purified fusion protein was obtained from 1L culture. Recombinant Mdmcec was readily obtained by enterokinase cleavage of the fusion protein followed by HPLC chromatography, and 11.2mg pure active recombinant Mdmcec was obtained from 1L E. coli culture. The molecular mass of recombinant Mdmcec determined by electrospray ionization-mass spectrometry (ESI-MS) is identical to that of native cecropin. Analysis of recombinant Mdmcec by circular dichroism (CD) indicated that recombinant Mdmcec contained predominantly alpha-helix with some random coil. Antimicrobial activity assays demonstrated that recombinant Mdmcec had a broad spectrum of activity against fungi, Gram-positive and negative bacteria. The procedure described in this study will provide a reliable and simple method for production of different cationic peptides for biological studies. 相似文献
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Isolation,purification, and N-terminal partial sequence of an antitumor peptide from the venom of the Chinese scorpion Buthus martensii Karsch 总被引:3,自引:0,他引:3
An antitumor peptide (ANTP) was isolated and purified from the venom of the Chinese scorpion Buthus martensii Karsch. The purification procedure included gel filtration on Sephadex G-50 and Superdex 30 high resolution chromatography, Phenyl Sepharose 6 Fast Flow chromatography, and SP-Sepharose Fast Flow chromatography. Its homogeneity was demonstrated by size exclusion HPLC on TSK G2000 SW. The isoelectric point is more than 10 by pH 3-10 range isoelectric focusing. ANTP has a relative molecular mass of 6280, calculated from the measurement of 16.5% SDS-PAGE. The pharmacological tests showed that ANTP has antitumoral effects in the mouse S-180 fibrosarcoma model and Ehrlich ascites tumor model. Amino acid analysis suggested the ANTP is rich in glycine and does not have histidine and threonine. The sequence of the first 25 N-terminal residues is as follows: Val-Arg-Asp-Gly-Tyr-Ile-Ala-Asp-Asp-Lys-Asn-Cys-Ala-Tyr-Phe-Cys-Gly-Arg-Asn-Ala-Tyr-Cys-Asp-Asp-Glu. 相似文献
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Bacterial infection poses an increasing threat to global public health and new types of antibacterial agents are urgently needed to respond to the threat. Scorpion venom contains series of bioactive peptides, among which antibacterial peptide is an important part. Herein, a new antimicrobial peptide StCT2 was characterized from the venomous gland cDNA library of the Scorpiops tibetanus. The full-length cDNA of StCT2 is 369 nucleotides encoding the precursor that contains a putative 24 residues signal peptide, a presumed 14 residues mature peptide, and a putative 37 residues acidic propeptide at the C-terminus. The minimal inhibition concentrations (MICs) of StCT2 for Staphylococcus aureus were 6.25-25μg/ml, including antibiotic-resistant strains such as methicillin resistant S. aureus (MRSA). StCT2 was further found to show high in vivo antimicrobial activity by an S. aureus infection mouse model. StCT2 exerted its antimicrobial activity via a rapid bactericidal mechanism. Taken together, these results demonstrate the efficacy and general mechanism of StCT2 antimicrobial action and the therapeutic potential of StCT2 as a new antimicrobial peptide. 相似文献
18.
The conditions for the sulfhydryl oxidase (SOX) production and activity from an Egyptian isolate of Aspergillus niger were optimized. Purification and determination of the kinetic properties (K
m and V
max) of the purified enzyme have been done. The possibility for the SOX induction using L-Cys (as a natural substrate) was studied
to determine whether SOX could be produced as an inducible enzyme in addition to being a constitutive one (i.e. whether induction
leads to increase SOX production and activity or not). The optimum temperature and pH for its activity were found to be 60°C
and 5.5, respectively. The activity of the induced intracellular SOX, was measured according to Ellman’s method using the
standard GSH oxidation where it reached 94% while that of non-induced one reached only 27.6%. This wide difference in activity
between the induced and non-induced SOX indicates the successful L—Cys-induction of the SOX production (i.e. SOX from A. niger AUMC 4947 is an inducible enzyme). Molecular characterization of the pure SOX revealed that it is constituted of two 50–55
KDa subunits. K
m and V
max were found to be 6.0 mM and 100 μM/min/mg respectively. 相似文献
19.
The conditions for the sulfhydryl oxidase (SOX) production and activity from an Egyptian isolate of Aspergillus niger were optimized. Purification and determination of the kinetic properties (K(m) and V(max)) of the purified enzyme have been done. The possibility for the SOX induction using L-Cys (as a natural substrate) was studied to determine whether SOX could be produced as an inducible enzyme in addition to being a constitutive one (i.e. whether induction leads to increase SOX production and activity or not). The optimum temperature and pH for its activity were found to be 60 degrees C and 5.5, respectively. The activity of the induced intracellular SOX, was measured according to Ellman's method using the standard GSH oxidation where it reached 94% while that of non-induced one reached only 27.6%. This wide difference in activity between the induced and non-induced SOX indicates the successful L-Cys-induction of the SOX production (i.e. SOX from A. niger AUMC 4947 is an inducible enzyme). Molecular characterization of the pure SOX revealed that it is constituted of two 50-55 KDa subunits. K(m) and V(max) were found to be 6.0 mM and 100 microM/min/mg respectively. 相似文献
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Panagiotou G Topakas E Economou L Kekos D Macris BJ Christakopoulos P 《Canadian journal of microbiology》2003,49(10):639-644
In the presence of L-arabinose as sole carbon source, Fusarium oxysporum produces two alpha-L-arabinofuranosidases (ABFs) named ABF1 and ABF2, with molecular masses of 200 and 180 kDa, respectively. The two F. oxysporum proteins have been purified to homogeneity. The purified enzymes are composed of three equal subunits and are neutral proteins with pIs of 6.0 and 7.3 for ABF1 and ABF2, respectively. With p-nitrophenyl alpha-L-arabinofuranoside (pNPA) as the substrate, ABF1 and ABF2 exhibited Km values of 0.39 and 0.28 mmol.L(-1), respectively, and Vmax values of 1.6 and 4.6 micromol.min(-1).(mg of protein)(-1), respectively, and displayed optimal activity at pH 6.0 and 50-60 degrees C. ABFs released arabinose only from sugar beet arabinan and not from wheat soluble and insoluble arabinoxylans. The enzymes were not active on substrates containing ferulic acid ester linked to C-5 and C-2 linkages of pNPA showing that phenolic substituents of pNPA sterically hindered the action of ABFs. 相似文献