一种新的从新疆穴居狼蛛中鉴定的细胞裂解肽

蜘蛛粗毒中富含生物活性物质,尤其以多肽类成分为主. 分离鉴定了新疆穴居狼蛛粗毒中一种新的细胞裂解肽,命名为LSTX-A1,其分子量为7 335.33,含有65个氨基酸残基,且羧基端残基酰胺化.研究了LSTX-A1的细胞裂解作用,结果显示,在100 μmol/L浓度引起42% 红细胞溶血.同时LSTX-A1具有抗肿瘤作用,能抑制HeLa细胞的增殖,其IC50剂量为22 μmol/L.     

新疆两种剧毒蜘蛛及其咬伤防治

介绍了新疆两种剧毒蜘蛛间斑寇蛛和穴居狼蛛的外形、生态,人被咬伤后的中毒症状以及防治方法。     

基于线粒体16S rRNA基因对中国狼蛛科(Lycosidae)系统发育研究

将自测的中国狼蛛科Lycosidae4亚科6属26种和从GenBank中检索到的北美2种豹蛛的mtD-NA-16S rRNA序列进行比较;以漏斗蛛科1种蜘蛛作为外群,对碱基序列的组成和遗传距离进行了分析,采用Bayesian方法和最大简约法(MP)构建分子系统树。研究结果表明:16SrRNA基因的部分序列为340bp到360bp,A T含量平均为75%,存在较强的A T含量偏向性;序列共有157个碱基存在变异,其中79个简约信息位点。狼蛛科各属间的遗传距离介于0.026￣0.200之间。2种建树方法均表明:科内的属及属内的种优先聚在一起;水狼蛛属相对马蛛属是狼蛛科中较为原始的类群,分化较早;獾蛛属作为1个单系群与熊蛛属合为1个并系,属于狼蛛亚科。狼蛛科6属间的分子系统关系为(Pirata(Hippasa(Trochsa Arctosa(Pardosa Wadicosa))))。     

狼蛛,蟹蛛和跳蛛

狼蛛、蟹蛛和跳蛛分别是蜘蛛目中三个科的蜘蛛的通称。这三类蜘蛛包括了我们日常见到的游猎型蜘蛛的大部分种类,它们在帮助人类消灭害虫中所起作用也较大,所以作为动物学教学中的例子加以介绍。 (一)狼蛛狼蛛在地面或植物上疾驰,凶狠如狼,故名。体长3-25毫米,但多数种类在5-8毫米间。体色多黄褐色,不鲜艳。8眼,排成三列。前列4个小眼,中、后两列各2个眼,较大;后列两眼的间距稍大于中列两眼的间距(图1左)。卵袋扁球形,由两片半圆形丝膜缝合而成。卵袋挂在母蛛腹部后端的纺器上,由母蛛随身携带。幼蛛孵出后不分散,而是爬伏在母蛛腹部     

中国狼蛛科娲蛛属(娲蛛亚科)分类地位的探讨

采用DNA测序方法,获得了中国狼蛛科Lycosidae4亚科6属26种mtDNA-16S rRNA基因的部分序列,比较来自北美狼蛛科豹蛛属2种豹蛛的同一基因序列,并选取漏斗蛛科1种蜘蛛作为外群,采用Bayesian方法和最大简约法(MP)构建分子系统树.两种建树方法均支持娲蛛属和豹蛛属形成一大的单系;这一结果与现行狼蛛科传统分类体系中娲蛛属的分类地位有差别.据此,作者认为:娲蛛属和豹蛛属可以归为同一个分类亚单位.狼蛛科6属间的分子系统关系为(Rirata(Hippasa(Trochsa Arctosa(Pardosa Wadicosa)))).     

虎纹捕鸟蛛毒素-III及其天然突变体的纯化与鉴定(英文)

虎纹捕鸟蛛毒素 III及其天然突变体是从虎纹捕鸟蛛粗毒中分离得到的两个毒素多肽。虎纹捕鸟蛛毒素 III含 33个氨基酸残基 ,其中包含 6个半胱氨酸残基 ;而其天然突变体只比虎纹捕鸟蛛毒素 III少了C端的色氨酸残基。MALDI TOF质谱测得虎纹捕鸟蛛毒素 III及其天然突变体的分子量分别为 385 3.35和 36 6 7.4 0。通过比较其理论分子量和质谱测定的分子量表明两个多肽的 6个半胱氨酸残基分别形成了三对二硫键。虎纹捕鸟蛛毒素 III与从同一种蜘蛛分离得到的凝集素 I具有 70 .5 %的序列相似性。生物学活性实验表明 ,虎纹捕鸟蛛毒素 III具有使美洲蜚蠊可逆的致瘫作用 ,其半有效剂量 (ED50 )为 (1 92 .95±1 2 0 .84 ) μg/g (P =0 .95 ) ,而且能加强由电刺激引起的大鼠输精管收缩 ;而其天然突变体却不具有上述生物学活性 ,表明C端色氨酸残基为虎纹捕鸟蛛毒素 III生物学活性相关残基 ;同时虎纹捕鸟蛛毒素 III及其天然突变体都不具有类似于凝集素 I对红细胞的凝集活性 ,表明虎纹捕鸟蛛毒素 III和凝集素 I两者氨基酸序列中不同氨基酸残基对于决定两者的生物学活性有着重要的作用     

我国狼蛛属1新种（蜘蛛目：狼蛛科）

报道采自湖北丹江的狼蛛1新种,即丹江狼蛛Lycosa danjiangensis sp.nov.。     

穴居狼蛛生活习性的初步观察

采用野外调查和室内、外饲养相结合的方法,观察研究了穴居狼蛛(Lycosa singoriensis)的捕食行为、繁殖习性、幼蛛生长发育、越冬、寿命等,并对成年雌蛛界定、互残行为及伤害人和动物等情况进行了讨论。     

中国狼逍遥蛛属1新种2新纪录种记述（蜘蛛目：逍遥蛛科）

记述分布于内蒙古的逍遥蛛科蜘蛛1新种:武川狼逍遥蛛Thannatus wuchuanensis sp.nov和2中国新纪录种:北极狼逍遥蛛Thanatus arctivus Thorell,1872;草原狼逍遥蛛Thanatus stepposus Logunov,1996.     

穴居狼蛛毒中磷酸单醌酶和核酸酶的研究

本文分别用对硝基酚磷酸酯、3-[32p]标记的RNA和大肠杆菌噬菌体入DNA作底物测定了新疆产穴居狼蛛毒中与核酸代谢有关的酶,发现此毒中含有磷酸单酯酶、核糖核酸酶和脱氧核糖核酸酶。     

Angiotensin I converting enzyme (ACE) inhibitory peptide derived from the sauce of fermented blue mussel, Mytilus edulis

The angiotensin I converting enzyme (ACE) inhibitory activity of fermented blue mussel sauce (FBMS) was investigated. Blue mussels were fermented with 25% NaCl (w/w) at 20 degrees C for 6 months and the resultant mixture was passed through a 40-mesh sieve, desalted using an electrodialyzer and then lyophilized. The IC(50) value of FBMS for ACE activity was 1.01 mg/ml. An ACE inhibitory peptide was purified from FBMS using Sephadex G-75 gel chromatography, SP-Sephadex C-25 ion exchange chromatography and reversed-phase high-performance liquid chromatography on a C(18) column. The IC(50) value of purified ACE inhibitory peptide was 19.34 microg/ml, and 10 amino acid residues of the N-terminal sequence was EVMAGNLYPG. The purified peptide was evaluated for antihypertensive effect in spontaneously hypertensive rats (SHR) following oral administration. Blood pressure significantly decreased after peptide ingestion. This result suggested that FBMS may have beneficial effects on hypertension.     

穴居狼蛛毒中抗菌活性物质的耐热及缔合性质

穴居狼蛛(Lycosa singoriensis)毒中的抗菌活性物质具有耐热性(在沸水浴中处理半小时活性不变)、耐酸性和耐碱性。但用透析袋(Visking 18/32)对毒腺提取物透析时,在中性及碱性条件下可全部被透析掉。当用Sephadex G-25柱对较少量样品进行层析时,在酸性条件下至少可被分出8个蛋白峰,其中峰4和峰8具抗菌活性,但随上柱样品量的增加,在层析图上峰4的面积增大,峰8的面积变小,甚至活性消失;反之,在中性或弱碱性条件下所得到的层析图上,则是峰8的面积大,抗菌活性明显,而峰4的面积小,甚至检不出抗菌活性。此峰8样品经热处理后,在酸性条件下经分子筛过滤,又可得到两个分子量不同的抗菌活性峰。这些结果提示,上述耐热和耐酸碱性强的抗菌物质在酸性环境中可能是以目身高聚体或与其它大分子蛋白非共价结合,随pH值的升高而解离成单体或低聚体。     

抗吗啡活性肽的分离纯化及一级结构测定

报道了一种新的具有抗吗啡镇痛活性的肽的分离纯化,并进行部分一级结构测定．狗脑先经醋酸提取,冷冻成干粉,然后上ＳｅｐｈａｄｅｘＧ－５０和Ｓ－ＳｅｐｈａｒｏｓｅＦ．Ｆ柱,最后经ＲＰ－ＨＰＬＣ纯化,鉴定纯度后,测定其抗吗啡镇痛活性,通过ＳＤＳ－ＰＡＧＥ法测得其分子量为８．９ｋＤ．氨基酸序列分析测得该肽的Ｎ端序列为：Ｖ－Ｉ－Ｓ－Ｖ－Ａ－Ｄ－Ｗ－Ｔ－Ｑ－Ｉ－Ｆ－Ｔ－Ｍ－Ｒ－Ｙ－Ｆ－Ｉ－Ｔ－Ｇ－Ｙ－Ｈ－Ｑ－Ｄ－Ｙ－Ｘ－Ｇ－Ｌ－Ｈ－Ｉ－Ｇ．经部分一级结构同源序列检索,未见与此有同源的蛋白质的报道,暂命名该肽为ＣＣ４肽     

Purification and partial characterization of lactacin F, a bacteriocin produced by Lactobacillus acidophilus 11088.

Lactacin F, a bacteriocin produced by Lactobacillus acidophilus 11088 (NCK88), was purified and characterized. Lactacin F is heat stable, proteinaceous, and inhibitory to other lactobacilli as well as Enterococcus faecalis. The bacteriocin was isolated as a floating pellet from culture supernatants brought to 35 to 40% saturation with ammonium sulfate. Native lactacin F was sized at approximately 180 kDa by gel filtration. Column fractions having lactacin F activity were examined by electron microscopy and contained micelle-like globular particles. Purification by ammonium sulfate precipitation, gel filtration, and high-performance liquid chromatography resulted in a 474-fold increase in specific activity of lactacin F. The purified bacteriocin was identified as a 2.5-kDa peptide by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The lactacin F peptide retained activity after extraction from SDS-PAGE gel slices, confirming the identity of the 2.5-kDa peptide. Variants of NCK88 that failed to exhibit lactacin F activity did not produce the 2.5-kDa band. Sequence analysis of purified lactacin F identified 25 N-terminal amino acids containing an arginine residue at the N terminus. Composition analysis indicates that lactacin F may contain as many as 56 amino acid residues.     

Purification and partial characterization of lactacin F, a bacteriocin produced by Lactobacillus acidophilus 11088

Lactacin F, a bacteriocin produced by Lactobacillus acidophilus 11088 (NCK88), was purified and characterized. Lactacin F is heat stable, proteinaceous, and inhibitory to other lactobacilli as well as Enterococcus faecalis. The bacteriocin was isolated as a floating pellet from culture supernatants brought to 35 to 40% saturation with ammonium sulfate. Native lactacin F was sized at approximately 180 kDa by gel filtration. Column fractions having lactacin F activity were examined by electron microscopy and contained micelle-like globular particles. Purification by ammonium sulfate precipitation, gel filtration, and high-performance liquid chromatography resulted in a 474-fold increase in specific activity of lactacin F. The purified bacteriocin was identified as a 2.5-kDa peptide by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The lactacin F peptide retained activity after extraction from SDS-PAGE gel slices, confirming the identity of the 2.5-kDa peptide. Variants of NCK88 that failed to exhibit lactacin F activity did not produce the 2.5-kDa band. Sequence analysis of purified lactacin F identified 25 N-terminal amino acids containing an arginine residue at the N terminus. Composition analysis indicates that lactacin F may contain as many as 56 amino acid residues.     

Purification and characterization of a bacteriocin-like compound (Lichenin) produced anaerobically by Bacillus licheniformis isolated from water buffalo

AIMS: To characterize a bacteriocin-like factor from Bacillus licheniformis 26 L-10/3RA isolated from buffalo rumen. METHODS AND RESULTS: The culture supernatant exhibited the antibacterial activity against a number of indicator organisms in a cut-well agar assay under anaerobic conditions. The inhibitory component was purified by following ammonium sulphate precipitation, gel filtration and ion exchange chromatography and confirmed to be a single peptide. A single band on tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis confirmed that the peptide was purified to homogeneity and having an estimated molecular mass of approximately 1400 dalton. Complete amino acid sequence of the peptide yielded 12 amino acids from the N-terminal end (ISLEICXIFHDN). No homology with previously reported bacteriocins was observed and has been designated as Lichenin. Lichenin was found to be hydrophobic, sensitive to atmospheric oxygen, retained biological activity even after boiling for 10 min and was active over a pH range of 4.0-9.0. CONCLUSIONS: The Lichenin represents the first anaerobiosis specific expression of bacteriocin-like compound isolated from Bacillus licheniformis 26 L-10/3RA of buffalo rumen origin. SIGNIFICANCE AND IMPACT OF THE STUDY: Lichenin could be a potential candidate for manipulating the rumen function at molecular level intended for improving the productivity of the ruminant.     

Processing of an antibacterial peptide from hemocyanin of the freshwater crayfish Pacifastacus leniusculus

An antibacterial peptide with 16 amino acid residues was found in plasma of the freshwater crayfish, Pacifastacus leniusculus. This peptide, designated astacidin 1, was purified by cation-exchange column chromatography and reverse-phase high performance liquid chromatography. Astacidin 1 has a broad range of antibacterial activity, and it inhibits growth of both Gram-positive and Gram-negative bacteria. The primary sequence of astacidin 1 was FKVQNQHGQVVKIFHH-COOH. The molecular mass was 1945.2 Da, and no carbohydrate-linked amino acid residues could be found by mass spectrometry. A synthetic astacidin 1 resulted in similar activity as the authentic astacidin 1 against Gram-positive bacteria, whereas it had less or no activity against Gram-negative bacteria. Three amino-terminal-truncated synthetic peptides were made; they all showed low activity, suggesting that the amino-terminal part of astacidin 1 contributes to the antibacterial activity. The structure of astacidin 1 based on the CD results showed that it has a beta-sheet structure in citric acid buffer at pH 4, 6, and 8. Cloning of astacidin 1 shows that it is the carboxyl-terminal part of crayfish hemocyanin and that astacidin 1 is produced by a proteolytic cleavage from hemocyanin under acidic conditions. The processing and release of astacidin 1 from hemocyanin is enhanced when crayfish are injected with lipopolysaccharide or glucan.     

类产碱假单胞菌杀虫蛋白的N端鉴定及抗体制备

The insecticidal protein from Pseudomonas pseudoaligenes was and exotoxin which had toxicity on locusts.In order to elucidate its molecular properties an amino acid sequence,the insecticidal protein was purified from the culture supernatant by ultrafiltration,ion\|exchange chromatography and gel filtration,and showed a single band on SDS\|PAGE.Analysis of the purified insecticidal protein dentified N\|terminal sequence of ten amino acid residues.Its polyclonal antibody was also obtained by immunizing rabb…     

Purification and characterization from human parotid secretion of a peptide which inhibits hemagglutination of Bacteroides gingivalis 381

A peptide from human parotid secretion which inhibited hemagglutination of Bacteroides gingivalis 381 was purified by ultrafiltration followed by DEAE-Sephadex A-25 column chromatography and by gel filtration on Sephadex G-25, and then by reversed-phase HPLC. The complete amino acid sequence of the peptide, determined by automated Edman degradation was as follows; Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr. The peptide contained 12 residues and the charged amino acids predominated with 4 histidine, 2 lysine, 1 arginine and 1 glutamic acid residues, thus being a histidine-rich peptide. The peptide was an active inhibitor of the hemagglutinating activity of B. gingivalis. Specific binding of tritium-labeled peptide to B. gingivalis cells was demonstrated. These results suggest that the histidine-rich peptide may function as a binding domain for the hemagglutinins of B. gingivalis during agglutination.     

Purification and cDNA cloning of rat 6-pyruvoyl-tetrahydropterin synthase

6-Pyruvoyl-tetrahydropterin synthase, which catalyzes the second step in the biosynthesis of tetrahydrobiopterin, was purified approximately 18,000-fold to apparent homogeneity from rat liver. The molecular mass of the native enzyme was estimated to be 83 kDa by gel filtration. The enzyme showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis corresponding to a molecular mass of 17 kDa. Up to 24 residues of the NH2-terminal sequence were determined by Edman degradation, which released a single amino acid at each step. These results indicate that the enzyme consists of identical subunits. The purified enzyme was digested with lysyl endopeptidase or V8 protease, and 11 peptide fragments were isolated. On the basis of the sequences of these peptides, oligonucleotides were synthesized and used to screen a rat liver cDNA library, and one cDNA clone was isolated. The complete nucleotide sequence of the 1176-base pair cDNA was then determined. The deduced amino acid sequence contained 144 amino acid residues, but a NH2-terminal four-amino acid sequence was not found in the purified protein. Therefore, the mature protein consists of 140 amino acids. A single mRNA band of 1.3 kilobases was obtained by RNA blot analysis of rat liver. The predicted amino acid sequence of 6-pyruvoyl-tetrahydropterin synthase was compared with the Protein Sequence Database of the National Biomedical Research Foundation, revealing significant local similarity to large T antigens from the polyomavirus family.