Two serine protease inhibitors from the skin secretions of the toad, Bombina microdeladigitora

Two serine protease inhibitors (named BMSI 1 and BMSI 2, respectively) were identified from the skin secretions of the toad, Bombina microdeladigitora. The cDNAs encoding BMSIs were cloned from a cDNA library prepared from the toad skin. The deduced complete amino acid sequences of BMSIs indicate that mature BMSI1 and BMSI2 are composed of 60 amino acids including 10 half-cystines to form 5 disulfide bridges. A FASTA search in the databanks revealed that BMSIs exhibit sequence similarity with other serine protease inhibitors from amphibians of the genus Bombina. BMSI1 potently inhibited trypsin and thrombin with a K(i) value of 0.02 mu M and 0.15 mu M, respectively. Sequence analysis revealed that all serine protease inhibitors from five amphibians of the genus Bombina share highly conserved primary structures. (c) 2007 Elsevier Inc. All rights reserved.

Molecular characterization of L-amino acid oxidase from king cobra venom

An L-amino acid oxidase from Ophiophagus hannah snake venom (Oh-LAAO) was purified by successive gel filtration, ion-exchange and heparin chromatography. Oh-LAAO did not induce platelet aggregation; however, it had potent inhibitory activity on platelet a

大蹼铃蟾缓激肽及其制备方法和其基因

本发明涉及大蹼铃蟾缓激肽及其制备方法和其基因，属于生物医学领域。其缓激肽为从中国两栖类动物大蹼铃蟾(Bombina maxima)皮肤分泌物中分离得到的由19个氨基酸组成的一种单链多肽，分子量2180，等电点11.7，多肽氨基酸全序列一级结构为：DLPKINRKGPRPPGFSPFR。制备方法是收集大蹼铃蟾皮肤分泌物，离心去除沉淀、冷冻干燥后，经凝胶过滤、高压液相反相柱层析分离纯化后即得到。编码大蹼铃蟾缓激肽基因核苷酸序列由cDNA由829个核苷酸组成，编码成熟大蹼铃蟾缓激肽为第140-196，224-280，308-364，392-448，476-532，560-616位6个核苷酸重复单元片断，从两栖类动物皮肤中得到的缓激肽基因作为基因工程制备缓激肽的应用。

A serine proteinase inhibitor from frog eggs with bacteriostatic activity

By Sephadex G-50 gel filtration, Resource Q anionic exchange and C4 reversed phase liquid high performance liquid chromatography, a proteinase inhibitor protein (Ranaserpin) was identified and purified from the eggs of the odour frog, Rana grahami. The protein displayed a single band adjacent to the molecular weight marker of 14.4 kDa analyzed by SDS-PAGE. The inhibitor protein homogeneity and its molecular weight were confirmed again by MALDI-TOF mass spectrometry analysis. The MALDI-TOF mass spectrum analysis gave this inhibitor protein an m/z of 14422.26 that was matched well with the result from SDS-PAGE. This protein is a serine proteinase inhibitor targeting multiple proteinases including trypsin, elastase, and subtilisin. Ranaserpin inhibited the proteolytic activities of trypsin, elastase, and subtilisin. It has an inhibitory constant (K-i) of 6.2 x 10(-8) M, 2.7 x 10(-7) M and 2.2 x 10(-8) M for trypsin, elastase, and subtilisin, respectively. This serine proteinase inhibitor exhibited bacteriostatic effect on Gram-positive bacteria Bacillus subtilis (ATCC 6633). It was suggested that ranaserpin might act as a defensive role in resistance to invasion of pests or pathogens. This is the first report of serine proteinase inhibitor and its direct defensive role from amphibian eggs. (C) 2007 Elsevier Inc. All rights reserved.

Identification and characterization of novel reptile cathelicidins from elapid snakes

Three cDNA sequences coding for elapid cathelicidins were cloned from constructed venom gland cDNA libraries of Naja atra, Bungarus fasciatus and Ophiophagus hannah. The open reading frames of the cloned elapid cathelicidins were all composed of 576 bp an

无指盘臭蛙胰岛素释放促进因子和在制药中的应用

本发明涉及无指盘臭蛙胰岛素释放促进因子和在制药中的应用，属于生物医学技术领域。本发明通过常规的生物化学手段，从无指盘臭蛙皮肤分泌液中分离纯化得到胰岛素释放促进因子肽并测定其序列，按照所得的序列合成该多肽。无指盘臭蛙胰岛素释放促进因子是一种单链多肽，分子量2648.4道尔顿，等电点 9.70，多肽全序列为：苯丙氨酸-亮氨酸-脯氨酸-亮氨酸-亮氨酸-丙氨酸- 甘氨酸-亮氨酸-丙氨酸-丙氨酸-天冬酰胺-苯丙氨酸-亮氨酸-脯氨酸- 赖氨酸-亮氨酸-苯丙氨酸-半胱苷酸-赖氨酸-异亮氨酸-苏氨酸-赖氨酸 -赖氨酸-半胱苷酸。本发明具有很好的促进胰岛素释放的作用，同时还具有无溶血活性、无血浆凝固活性等优点，可作为制备治疗糖尿病药物的应用。

Toward an understanding of the molecular mechanism for successful blood feeding by coupling proteomics analysis with pharmacological testing of horsefly salivary glands

Horseflies are economically important blood-feeding arthropods and also a nuisance for humans and vectors for filariasis. They rely heavily on the pharmacological properties of their saliva to get a blood meal and suppress immune reactions of hosts. Little information is available on antihemostatic substances in horsefly salivary glands; especially no horsefly immune suppressants have been reported. By proteomics or peptidomics and coupling transcriptome analysis with pharmacological testing, several families of proteins or peptides, which act mainly on the hemostatic system or immune system of the host, were identified and characterized from 30,000 pairs salivary glands of the horsefly Tabanus yao (Diptera, Tabanidae). They are: (i) a novel family of inhibitors of platelet aggregation including two members, which possibly inhibit platelet aggregation by a novel mechanism and act on platelet membrane, (ii) a novel family of immunosuppressant peptides including 12 members, which can inhibit interferon-gamma production and increase interleukin-10 secretion, (iii) a serine protease inhibitor with 56 amino acid residues containing anticoagulant activity, (iv) a serine protease with anticoagulant activity, (v) a protease with fibrinogenolytic activity, (vi) three families of antimicrobial peptides including six members, (vii) a hyaluronidase, (viii) a vasodilator peptide, which is an isoform of vasotab identified from Hybomitra bimaculata, and interestingly (ix) two metallothioneins, which are the first metallothioneins reported from invertebrate salivary glands. The current work will facilitate the understanding of the molecular mechanisms of the ectoparasite-host relationship and help in identifying novel vaccine targets and novel leading pharmacological compounds.

Isolation, expression and characterization of a novel dual serine protease inhibitor, OH-TCI, from king cobra venom

Snake venom Kunitz/BPTI members are good tools for understanding of structure-functional relationship between serine proteases and their inhibitors. A novel dual Kunitz/BPTI serine proteinase inhibitor named OH-TCI (trypsin- and chymotrypsin-dual inhibito

无指盘臭蛙蛙皮肽及其基因和在制药中的应用

本发明涉及一种无指盘臭蛙蛙皮肽及其基因和在制药中的应用，属于生物医学领域。该活性多肽是中国两栖类动物无指盘臭蛙基因编码的一种单链多肽，分子量3355.95道尔顿，等电点10.14，多肽全序列一级结构为： Gly Leu Gly Gly Ala Lys Lys Asn Phe Ile Ile Ala Ala Asn Lys Thr Ala Pro Gln Ser Val Lys Lys Thr Phe Ser Cys Lys Leu Tyr Asn Gly。编码无指盘臭蛙蛙皮肽的基因由 731个核苷酸组成，其中编码成熟无指盘臭蛙蛙皮肽为第520-616位核苷酸。人工合成的无指盘臭蛙蛙皮肽具有显著的抑制细菌和真菌生长的作用，可以作为制备病原微生物感染疾病的治疗药物被应用。本发明的无指盘臭蛙蛙皮肽具有结构简单、人工合成方便、抗菌谱系广的有益特点。

A phospholipase A1 platelet activator from the wasp venom of Vespa magnifica (Smith)

Wasp is an impor tant venomous animal that can induce human fatalities. Aortic thrombosis and cerebral infarction are major clinical symptoms after massive wasp stings but the reason leading to the envenomation manifestation is still not known. In this paper, a toxin protein is purified and characterized by Sephadex G-75 gel filtration, CM-Sephadex C-25 cationic exchange and fast protein liquid chromatography (FPLC) from the venom of the wasp, Vespa magnifica (Smith). This protein, named magnifin, contains phospholipase-like activity and induces platelet aggregation. The cDNA encoding magnifin is cloned from the venom sac cDNA library of the wasp. The predicted protein was deduced from the cDNA with a sequence composed of 337 amino acid residues. Magnifin is very similar to other phospholipase A(1) (PLA(1)), especially to other wasp allergen PLA(1). Magnifin can activate platelet aggregation and induce thrombosis in vivo. The current results proved that PLA(1) in wasp venom could be contributable to aortic thrombosis after massive wasp stings. (c) 2007 Elsevier Ltd. All rights reserved.

beta gamma-CAT, a non-lens beta gamma-crystallin and trefoil factor complex from amphibian skin secretions, caused endothelium-dependent myocardial depression in isolated rabbit hearts

Previous in vivo study demonstrated that beta gamma-CAT, a newly identified non-lens beta gamma-crystallin and trefoil factor complex from frog Bombina maxima skin secretions, possessed potent lethal toxicity on mammals resulted from hypotension and cardi

无指盘臭蛙皮肤活性肽及其基因和在制药中的应用

本发明涉及一种无指盘臭蛙皮肤活性肽及及其基因和在制药中的应用，属于生物医学领域。无指盘臭蛙皮肤活性肽是无指盘臭蛙皮肤活性肽基因编码的一种单链多肽，分子量3015.63道尔顿，等电点10.3，多肽全序列一级结构为： Ala Thr Ala Leu Gly Leu Ser Ser Arg Gly Leu Leu Pro Ile Gly Phe Met Phe Lys Asp Thr Ile Arg Cys Arg Lys Tyr(ATALGLSSRGLLPIGFMFKDTIRCRKY)。编码无指盘臭蛙皮肤活性肽的基因由312个核苷酸组成，其中编码成熟无指盘臭蛙皮肤活性肽为第141-222位核苷酸。人工合成的无指盘臭蛙皮肤活性肽具有很强的抑制细菌和真菌生长的作用，可以作为制备病原微生物感染疾病的治疗药物被应用。无指盘臭蛙皮肤活性肽具有结构简单、人工合成方便、抗菌谱系广、抗菌活性强的特点。

The first antimicrobial peptide from sea amphibian

The crab-eating frog, Rana cancrivora, is one of only a handful of amphibians worldwide that tolerates saline waters. It typically inhabits brackish water of mangrove forests of Southeast Asia. A large amount of antimicrobial peptides belonging to different families have been identified from skins of amphibians inhabiting freshwater. No antimicrobial peptide from sea amphibians has been reported. In this paper, we firstly reported the antimicrobial peptide and its cDNA cloning from skin secretions of the crab-eating frog R. cancrivora. The antimicrobial peptide was named cancrin with an amino acid sequence of GSAQPYKQLHKVVNWDPYG. By BLAST search, cancrin had no significant similarity to any known peptides. The cDNA encoding cancrin was cloned from the cDNA library of the skin of R. cancrivora. The cancrin precursor is composed of 68 amino acid residues including a signal peptide, acidic spacer peptide, which are similar to other antimicrobial peptide precursors from Ranid amphibians and mature cancrin. The overall structure is similar to other amphibian antimicrobial peptide precursors although mature cancrin is different from known peptides. The current results reported a new family of amphibian antimicrobial peptide and the first antimicrobial peptide from sea amphibian. (c) 2007 Elsevier Ltd. All rights reserved.

Melanoma cell growth inhibition by beta gamma-CAT, which is a novel non-lens betagamma-crystallin and trefoil factor complex from frog Bombina maxima skin

In vertebrates, non-lens beta gamma-crystallins are widely expressed in various tissues but their functions are unknown. The molecular mechanisms of trefoil factors, initiators of mucosal healing and being greatly involved in tumorigenesis, have remained

无指盘臭蛙免疫调节肽、其基因和变异体及其在制药中的应用

本发明涉及无指盘臭蛙免疫调节肽、基因和变异体及其在制药中的应用，属生物医学技术领域。无指盘臭蛙免疫调节肽是一种环状多肽，分子量1791.12道尔顿，等电点 9.84，无指盘臭蛙免疫调节肽的全序列为：Thr Ser Arg Cys Tyr Ile Gly Tyr Arg Arg Lys Val Val Cys Ser(TSRCYIGYRRKVVCS)，其第4位的半胱氨酸和第14位的半胱氨酸形成分子内二硫键。编码的基因由307个核苷酸组成，其中编码成熟部分的为第 141-186位核苷酸。无指盘臭蛙免疫调节肽原始序列中第4个氨基酸发生替代所产生的变异体，人工合成的无指盘臭蛙免疫调节肽及其变异体具有强烈的免疫调节活性和肿瘤抑制活性，作为制备免疫调节、肿瘤治疗和化疗药物的应用。本发明还具有序列简单、合成方便等优点。