Cloning of novel bombesin precursor cDNAs from skin of Bombina maxima

Amphibian skin is a rich resource of bioactive peptides like proline-rich bombesin from frog Bombina maxima. A novel cDNA clone encoding a precursor protein that comprises proline-rich bombesin and a novel peptide, designated as bombestatin, was isolated from a skin cDNA library of B. maxima. The predicted primary structure of the novel peptide is WEVLLNVALIRLELLSCRSSKDQDQKESCGMHSW, in which two cysteines form a disulfide bond. A BLAST search of databases did not detect sequences with significant similarity. Bombestatin possesses dose-dependent contractile activity on rat stomach strips. The differences between cDNAs encoding PR-bombesin plus bombestatin and PR-bombesin alone are due to fragment insertions located in 3'-coding region and 3'-untranslational region, respectively. (c) 2005 Elsevier B.V. All rights reserved.

Vespid chemotactic peptide precursor from the wasp, Vespa magnifica (Smith)

Despite the evolutional distance between wasp and amphibian, vespid chemotactic peptide (VCP), an important component of wasp venom, are found sharing remarkable similarities with the temporin antimicrobial peptides (AMPs) from Ranid frog, Amolops loloens

蹼铃蟾蛋白酶抑制剂及其制备方法和在制药中的应用

本发明属于生物医学领域。大蹼铃蟾蛋白酶抑制剂，从两栖类动物大蹼铃蟾皮肤中分离得到的一种单链丝氨酸蛋白酶抑制剂，其在SDS－聚丙烯酰胺凝胶电泳上的表观分子量还原条件下为67KDa，非还原条件下为45KDa，等电点4.8，糖含量17—19％，无酶活性，大蹼铃蟾蛋白酶抑制剂的N—端31个氨基酸序列结构是：DSETHIRFLGEVYKKVDTIDFRGLVLITRAQ。其制备方法是取大蹼铃蟾皮肤，在生理盐水中匀浆后，离心去除沉淀，收集上清液冷冻干燥后，经离子交换，凝胶过滤纯化后得到。具有强烈的丝氨酸蛋白酶抑制活性和抗肿瘤活性，作为制备肿瘤治疗药物和制备治疗胃炎、胰腺炎及其他炎症药物的应用。

A novel heme-containing protein with anti-HIV-1 activity from skin secretions of Bufo andrewsi

A novel protein, named BAS-AH, was purified and characterized from the skin of the toad Bufo andrewsi. BAS-AH is a single chain protein and the apparent molecular weight is about 63 kDa as judged by SDS-PAGE. BAS-AH was determined to bind heme (0.89 mol heme/mol protein) as determined by pyridine haemochrome analysis. Fifty percentage cytotoxic concentration (CC50) of BAS-AH on C8166 cells was 9.5 mu M. However, at concentrations that showed little effect oil cell viability, BAS-AH displayed dose dependent inhibition oil HIV-1 infection and replication. The antiviral selectivity indexes corresponding to the measurements of syncytium formation and HIV-1 p24 (CC50/EC50) were 14.4 and 11.4, respectively, corresponding to the . BAS-AH also showed an inhibitory effect on the activity of recombinant HIV-1 reverse transcriptase (IC50 = 1.32 mu M). The N-terminal sequence of BAS-AH was determined to be NAKXKADVIGKISILLGQDNLSNIVAM, which exhibited little identity with other known anti-HIV-1 proteins. BAS-AH is devoid of antibacterial, protcolytic, trypsin inhibitory activity, (L)-amino acid oxidase activity and catalase activity. (c) 2005 Elsevier Ltd. All rights reserved.

Trypsin inhibitory loop is an excellent lead structure to design serine protease inhibitors and antimicrobial peptides

The disulfide-bridged hendecapeptide ( CWTKSIPPKPC) loop, derived from an amphibian skin peptide, is found to have strong trypsin inhibitory capability. This loop, called the trypsin inhibitory loop ( TIL), appears to be the smallest serine protease inhib

大蹼铃蟾抗精子多肽及其制备方法和在制药中的应用

大蹼铃蟾抗精子多肽及其制备方法和在制药中的应用，属生物医学领域。大蹼铃蟾抗精子多肽，其在聚丙烯酰胺凝胶多肽电泳上的表观分子量还原和非还原条件下为8000Da，等电点8.8；其N－端20个氨基酸残基序列是：AVITGVICRRAQCGVGCAER。制备方法是将活体大蹼铃蟾清洗干净，经收集皮肤分泌物、离心去除沉淀、冷冻干燥、离子交换、凝胶过滤纯化等工序后得到。大蹼铃蟾抗精子多肽具有较强的精子制动活性和丝氨酸蛋白酶抑制活性，作为制备避孕药物和制备治疗胰腺炎及其他炎症药物的应用。

Purification and characterization of an irreversible serine protease inhibitor from skin secretions of Bufo andrewsi

Amphibian skin secretions contain many bioactive compounds. In the present work, an irreversible serine protease inhibitor, termed baserpin, was purified for the first time from the skin secretions of toad Bufo andrewsi by Successive ion-exchange and gelfiltration chromatography. Baserpin is a single chain glycoprotein, with an apparent molecular weight of about 60 kDa in SDS-PAGE. Baserpin is an irreversible inhibitor and effectively inhibits the catalytic activity of trypsin, chymotrypsin and elastase. SDS-stable baserpin-trypsin complex could be seen in SDS-PAGE indicates that it possibly belongs to the serpin superfamily. According to the association rates determined, baserpin is a potent inhibitor of bovine trypsin (4.6 X 10(6) M-1 S-1), bovine chymotrypsin (8.9 X 10(6) M-1 s(-1)) and porcine elastase (6.8 X 10(6) M-1 s(-1)), whereas it shows no inhibitory effect on thrombin. The N-terminal sequence of baserpin is HTQYPDILIAKPXDK, which shows no similarity with other known serine protease inhibitors. (c) 2005 Elsevier Ltd. All rights reserved.

A bactericidal homodimeric phospholipases A(2) from Bungarus fasciatus venom

Group IIA secretory phospholipases A(2) (sPLA(2)-II) is generally known to display potent grampositive bactericidal activity, while group IA sPLA(2) (sPLA(2)-I) reportedly is not. In this work, a novel sPLA(2)-I named BFPA was identified from Bungarus fas

大蹼铃蟾抗肿瘤蛋白及其制备方法和其基因

本发明涉及大蹼铃蟾抗肿瘤蛋白及其制备方法和其基因，属于生物医学领域。抗肿瘤蛋白是从中国两栖类动物大蹼铃蟾皮肤分泌物中分离得到的一种单链多肽，分子量2697，等电点9.83，多肽氨基酸全序列一级结构为：NH2－GIGGKILSGLKTALKGAAKELASTYLH－NH2。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，经离子交换、高压液相反相柱层析分离纯化后得到。由650、623、625个核苷酸组成的三个不同的基因可编码大蹼铃蟾抗肿瘤蛋白，编码成熟抗肿瘤蛋白分别为第177－257位、第166－246位、175－255位核苷酸。具有抑制肿瘤细胞生长、抗艾滋病病毒活性和抑制细菌和真菌生长的作用。

Frog albumin is expressed in skin and characterized as a novel potent trypsin inhibitor

A novel potent trypsin inhibitor was purified and characterized from frog Bombina maxima skin. A full-length cDNA encoding the protein was obtained from a cDNA library constructed from the skin. Sequence analysis established that the protein actually comprises three conserved albumin domains. B. maxima serum albumin was subsequently purified, and its coding cDNA was further obtained by PCR-based cloning from the frog liver. Only two amino acid variations were found in the albumin sequences from the skin and the serum. However, the skin protein is distinct from the serum protein by binding of a haem b (0.95 mol/mol protein). Different from bovine serum albumin, B. maxima albumin potently inhibited trypsin. It bound tightly with trypsin in a 1: 1 molar ratio. The equilibrium dissociation constants (K-D) obtained for the skin and the serum proteins were 1.92 x 10(-9) M and 1.55 x 10(-9) M, respectively. B. maxima albumin formed a noncovalent complex with trypsin through an exposed loop formed by a disulfide bond (Cys(53)-Cys(62)), which comprises the scissile bond Arg(58)(P-1)-His(59)(P-1'). No inhibitory effects on thrombin, chymotrypsin, elastase, and subtilisin were observed under the assay conditions. Immunohistochemical study showed that B. maxima albumin is widely distributed around the membranes of epithelial layer cells and within the stratum spongiosum of dermis in the skin, suggesting that it plays important roles in skin physiological functions, such as water economy, metabolite exchange, and osmoregulation.

Anti-infection peptidomics of amphibian skin

Peptidomics and genomics analyses were used to study an anti-infection array of peptides of amphibian skin. 372 cDNA sequences of antimicrobial peptides were characterized from a single individual skin of the frog Odorrana grahami that encode 107 novel an

大蹼铃蟾血管扩张因子及其制备方法和其基因

本发明涉及大蹼铃蟾血管扩张因子及其制备方法和其基因，属于生物医学领域。其血管扩张因子为从中国两栖类动物大蹼铃蟾(Bombinamaxima)皮肤分泌物中分离得到的一种单链多肽，分子量2985，等电点10.2，多肽氨基酸全序列一级结构为：NH2－GIGRKFLGGVKTTFRCGVKDFASKHLY－NH2。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，经离子交换、高压液相反相柱层析分离纯化后即得到。编码血管扩张因子的基因由647个核苷酸组成，其中编码成熟血管扩张因子为第178－258位核苷酸。其血管扩张因子具有显著的扩张血管特别是微血管的作用，其基因作为基因工程制备血管扩张因子的应用。

Isolation and preliminary characterization of a 22-kDa protein with trypsin inhibitory activity from toad Bufo andrewsi skin

A novel trypsin inhibitor termed BATI was purified to homogeneity from the skin extracts of toad Bufo andrewsi by successive ion-exchange, gel-filtration and reverse-phase chromatography. BATI is basic single chain glycoprotein, with apparent molecular weight of 22 kDa in SDS-PAGE. BATI is a thermal stable competitive inhibitor and effectively inhibits trypsin's catalytic activity on peptide substrate with the inhibitor constant (K-i) value of 14 nM and shows no inhibitory effect on chymotrypsin, thrombin and elastase. The N-terminal sequence of BATI is EKDSITD, which shows no similarity with other known trypsin inhibitors. (c) 2005 Elsevier Ltd. All rights reserved.

Pontibacter akesuensis sp nov., isolated from a desert soil in China

A Gram-negative, non-motile, rod-shaped bacterium, designated strain AKS 1 T, was isolated from a desert soil sample collected from Alkesu, Xin.lang Province, China. A taxonomic study, including phylogenetic analysis based on 16S rRNA gene sequences and p

大蹼铃蟾抗微生物肽及其制备方法和其基因

本发明涉及一种大蹼铃蟾抗微生物肽及其制备方法和其基因，属于生物医学领域。抗微生物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一种单链多肽，分子量2840，等电点983，多肽氨基酸全序列一级结构为：NH2－SIGAKILGGVKTFFKGALKELASTYLQ－NH2。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，经离子交换、高压液相反相柱层析分离纯化后得到。编码抗微生物肽的基因由663个核苷酸组成，其中编码成熟抗微生物肽为第165－245位核苷酸。抗微生物肽具有广谱和显著的抑制细菌和真菌生长作用。抗微生物肽基因作为基因工程制备抗微生物肽的应用。