Analysis of the hemorrhagic principle of the venom of Echis multisquamatus

Institute of Biochemistry, Uzbek SSR Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 438–439, May–June, 1989.     

一种新的蛇毒解离素同系物的分离

为发现新的具有生物活性的蛇毒蛋白,以我国华南地区富有的蝮蛇蛇毒为分离样品源,采用高效液相色谱,从中分离出一种新的解离素同系物———AHP-4,建立了快速分离纯化蛇毒蛋白的方法。结构确认的结果表明,AHP-4的相对分子质量为7 554 Da,是由一条多肽链组成的蛋白质,N-端第1~15位的氨基酸序列为GEECDCGSPANPCCD。经GeneBank protein-protein BLAST检索及人工检索,在已知蛋白质N-端第1~15位氨基酸序列中,未发现与AHP-4氨基酸结构完全相同的蛋白质。AHP-4 N端第1~5位氨基酸序列与5种解离素中的氨基酸序列具有高度同源性,表明AHP-4是一种新的解离素同系物。     

三步柱色谱法从江浙蝮蛇蛇毒中分离纯化类凝血酶

建立了一种用CM Sepharose CL-6B阳离子交换、DEAE Sepharose Fast Flow阴离子交换和Sephadex G-75凝胶排阻三步柱色谱从江浙蝮蛇蛇毒中分离纯化类凝血酶的方法。在实验室小柱分离方案的基础上,对该纯化工艺进行了放大。当上样量达实验室小柱的25倍时,所得类凝血酶的质量指标与实验室小柱基本一致。采用该法所得的蝮蛇类凝血酶经Shim-pack Diol-300高效凝胶排阻柱测得其相对分子质量约为33500,用Shim-pack VP-ODS反相色谱柱检测其纯度约为96%。从江浙粗蛇毒中提取类凝血酶时,类凝血酶的总质量收率约为0.3%,总活性收率约为64%,比活可达2000 U/mg。     

Preparation of a Monoclonal Antibody against a Kallikrein-Like Enzyme from Agkistrodon halys pallas Venom and Its Application in a Pharmacokinetic Study

Snake venom contains bioactive materials for drug development, diagnosis, and treatment. After separating and purifying the kallikrein-like enzyme (AHP-Ka) from Agkistrodon halys pallas venom for the first time, a monoclonal antibody against AHP-Ka was prepared and characterized. An indirect sandwich enzyme-linked immunosorbent assay (ELISA) based on the monoclonal antibody was developed and validated for the pharmacokinetic analysis of AHP-Ka in rat plasma. The method was calibrated using rat plasma and 1:100 dilution of plasma was selected to prepare a calibration curve to validate the precision, accuracy, and stability of the ELISA method. A good linear relationship was obtained in a working range from 3.9 ng/mL to 62.5 ng/mL with a limit of detection of 2.94 ng/mL. Intra- and inter-batch precision were less than 10%. The average recovery ranged from 94.6% to 104.4% in rat plasma at the concentrations of 5 ng/mL, 15 ng/mL, and 45 ng/mL, respectively. The ELISA method was successfully used for the pharmacokinetic study of AHP-Ka in Sprague-Dawley rat plasma after intravenous administration. The work is expected to contribute to future preclinical development of AHP-Ka.     

Antioxidant activities of extracts and fractions from Eupatorium lindleyanum DC

The antioxidant activities of water extract (WE), ethanol extract (EE), residue water extract (RWE) and petroleum ether (PF), ethyl acetate (EF), n-BuOH (BF) and water (WF) fractions of the ethanol extract from Eupatorium Lindley DC were investigated for the first time. Total phenolics content, DPPH radical scavenging activities, superoxide radical scavenging activities, total reduction capability, and ferrous ions chelating activities were determined for all the extracts and fractions. The results showed that all the extracts and fractions exhibited antioxidant activities with different magnitudes of potency. Among all the samples, WE and RWE exhibited the best antioxidant capacities, the BF also exhibited high antioxidant abilities in all tests except for the metal chelating activity, while the other extracts and fractions were relatively weak antioxidants. The BF had the highest total phenolics contents in all extracts and fractions, and the WE and RWE were found to be rich in tannins. Furthermore, the content of total phenolics showed good correlation with DPPH radical scavenging activity, superoxide anion radical scavenging activity, and the reducing power. Phenolic composition of all the extracts and fractions was identified and quantified by HPLC. The results indicate that the extracts of E. Lindley DC might be a useful potential source of natural antioxidant ingredients.     

Inhibition of angiotensin-converting enzyme by peptides from the venom of the viperEchis multisquamatus

Angiotensin I-converting enzyme [dipeptidyl carboxypeptidase] (ACE, CE 3.4.15.1) isolated in the pure form from ox kidney, has been characterized physicochemically and kinetically. Inhibition of the enzyme by homogeneous peptides from the venom of the Central Asian viperEchis multisquamatus Ch. has been investigated. Institute of Biochemistry, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (371) 62 32 56. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 821–825, November–December, 1998.     

Peptide inhibitors of angiotensin I-converting enzyme from the venom ofEchis multisquamatus

Using bradykinin (BK) as substrate and ACE from ox kidneys we have studied the inhibitory properties of eight peptides isolated in the pure form from viper venom. The values of IC-50 for them proved to be between 1.6 and 265.0 g/ml. It was shown simultaneously that the inhibitors isolated, consisting of hydrophobic positively charged deca- to pentapeptides, potentiate the contractile activity of BK on smooth-muscle preparations in vitro. For a set of six of the peptides, practically complete coincidence of a number of potentiating activites and inhibitory properties has been demonstrated (correlation coefficient r = 0.86 ± 0.14). The inclusion of the two other peptides in the analysis leads to a markedfall in the corrrelation and shows the existence in their case of additional mechanisms for the realization of bradykinin potentiating action.Institute of Biochemistry, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (3712) 62 32 56. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 441–444, May–June, 1995. Original article submitted October 17, 1994.     

Competitive inhibitors of the angiotensinconverting enzyme from the venom of the viperEchis multisquamatus

The mechanism of the inhibition of the angiotensin-converting enzyme (ACE, E. C. 3.4.15.1) by homogeneous peptides of the venom of the Central Asian viperEchis multisquamatus Ch. has been determined, and the inhibition constants have been measured, by means of an analysis of Lineweaver-Burk graphs.Institute of Biochemistry, Academy of Sciences of the Republic of Uzbekistan, Tashkent fax (3712) 62 32 56. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 368–371, May–June, 1999.     

Physicochemical analysis of a protein with nerve growth activity from the muscle tissue of Rana ridibunda

A protein nerve growth factor has been isolated in the pure form from the muscles of the lake frogRana ridibunda. The individuality of the substance obtained has been shown, its molecular mass (106.0–110.0 kDa) and pI value (9.2–9.4) have been determined, and lysine and valine have been found as the N- and C-terminal amino acid residues of its polypeptide chain. In view of the detection of the presence of sugars (about 1%), the substance has been identified as a glycoprotein. Its absorption spectra in the UV region has been obtained and the amino acid composition of the isolated protein has been determined. Information on the amino acid composition of various proteins with nerve growth activity has been subjected to comparative analysis with the aid of computer programs, and on this base their phylogenetic tree has been constructed.Institute of Biochemistry, Uzbekistan Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 251–257, March–April, 1992.     

Mitogenic and colony-stimulating factor-inducing activities of polysaccharide fractions from the fruit bodies of Dictyophora indusiata Fisch

Biological effects (mitogenic and colony-stimulating factor (CSF)-inducing activities) of five homogeneous polysaccharides and a conjugated polysaccharide fraction isolated from the fruit bodies of Dictyophora indusiata Fisch. were investigated. Fucomammogalactan (T-3-Ad) and conjugated polysaccharide fraction (T-2-A) exhibited significant mitogenic and CSF-inducing activities. Among two beta-(1----6)-branched (1----3)-beta-D-glucans (T-4-N and T-5-N), only T-4-N showed both mitogenic and CSF-inducing effects. Partially O-acetylated (1----3)-alpha-D-mannans (T-2-HN and T-3-M') did not show these effects.     

毛细管区带电泳分析五步蛇毒蛋白C激活剂和蛋白C的相互作用

建立了毛细管区带电泳分析五步蛇毒蛋白C激活剂(protein C activator, PCA)与血浆蛋白C(protein C, PC)的相互作用。以未涂层毛细管(60.2 cm(有效长度50 cm)×75 μm)为分离柱,50 mmol/L Tris-HCl(pH 7.4)为运行缓冲液,于198 nm波长下检测。对影响五步蛇毒PCA分离的因素(如缓冲液、离子浓度)及在37.5 ℃下孵育不同时间的五步蛇毒PCA与PC的相互作用进行考察。方法的检出限(以信噪比为3计)为3 mg/L,线性范围为10～300 mg/L。五步蛇毒PCA迁移时间和峰面积的相对标准偏差分别为0.56%和3.8%(n=6)。等体积的五步蛇毒PCA(200 mg/L)与PC(60 mg/L)孵育5 min,其结合率达到最大,且谱图中未见有PC水解的肽链。该五步蛇毒PCA可改变PC空间构象直接激活PC。该方法简单,灵敏度和分辨率高,分析结果为今后快速检测五步蛇毒PCA及其活性提供了重要的理论依据。     

Features of the circular dichroism spectrum of protein fractions from opaque-2 maize

Scientific-Research Institute of Biology, Dnepropetrovsk State University. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 131–132, January–February, 1990.     

长白山白眉蝮蛇蛇毒磷脂酶A2的荧光光谱

利用荧光光谱系统研究了长白山白眉蝮蛇蛇毒磷脂酶Ａ２的荧光特性。研究结果表明：当发射波长与激发波长差为２０ｎｍ和７５ｎｍ时,ＰＬＡ２的同步荧光分别主要由酷氨酸（Ｔｙｒ）残基和色氨酸（Ｔｒｐ）残基所贡献。缓冲溶液的酸度变化能够明显影响ＰＬＡ２氨基酸侧链的电荷分布,从而改变ＰＬＡ２的荧光发射强度。了子Ｃａ＾２＋一方面能增强ＰＬＡ２的荧光强度,另一方面也能够加快ＰＬＡ２与底物二棕樟酰磷脂酰胆碱（ＤＰＰＣ）     

Heterogeneity in the conformation of different protein fractions from the human erythrocyte membrane.

We have isolated 5 families of proteins from human red blood cell membranes and characterized their secondary structure by ultraviolet circular dichroism measurements. The protein families were prepared by selective solubilization from ghosts under nondenaturing conditions. We find that the intact ghost has a mean alpha-helix fraction of 0.37, whereas a low-ionic-strength extract (bands 1, 2, 5, "spectrin") has a substantially higher helix fraction, 0.55. Further extraction of the ghosts with para-chloromercuribenzoate yields bands 2.1, 4.1, 4.2, and 6; their helix content is only 0.17. Finally, the major intrinsic protein, band 3, was solubilized by a non-ionic detergent. Its helix fraction is 0.38.     

Phosphatase activities of fractions of the venom of Renard's viperVipera ursini renardi

The separation of the venom of Renard's viper by filtration through Sephadex G-75 gel has been performed and phosphatase activities have been determined in the fractions and in the course of separation. The bulk of the activity of the phosphatases investigated was concentrated in the high-molecular-weight fractions I and II. A correlation has been found in the distribution of the DNase and ATP-pyrophosphatase activities with the phosphodiesterase activity in the venom fractions. There is no correlation in the distribution of the RNase and phosphodiesterase activity.     

Some properties of the protein fractions of tomato seeds

The solubilities and rheological properties of interphase adsorption layers (IALs) of the protein fractions of tomato seeds have been studied. Regions of minimum solubility have been determined. It has been established that the globulin fraction exhibits pronounced surface properties, forming strong IALs at liquid phase-separation boundaries. A correlation has been found between the rheology of IALs and the stability of emulsions of the oil-in-water type. Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (3712) 62 73 48. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 537–540, July–August, 1998.