Monteiro R Q, Dutra D L, Machado O L, Carlini C R, Guimarães J A, Bon C, Zingali R B
Departamento de Bioquímica Médica, ICB-CCS-UFRJ, Cidade Universitária, Rio de Janeiro, Brazil.
Comp Biochem Physiol B Biochem Mol Biol. 1998 Aug;120(4):791-8. doi: 10.1016/s0305-0491(98)10070-6.
More than one isoform of bothrojaracin (BJC), a potent and specific thrombin inhibitor isolated from Bothrops jararaca venom, has been found in individual venoms collected from adult snakes. Variations in snake venom composition have previously been associated with factors such as age, sex, geographic origin, season of the year and diet. In order to obtain further information concerning individual patterns of expression of BJC isoforms, we have analyzed five individual Bothrops jararaca snake venoms collected at the same time from adult female snakes from the same geographic region. As expected, crude venoms showed a similar migration pattern on SDS-PAGE. BJC was purified using a procedure which includes an affinity chromatography step (PPACK-thrombin Sepharose). A slight variation in the amount of BJC obtained from individual venom samples was noticed. Inhibition of thrombin-induced platelet aggregation as well as migration pattern on SDS-PAGE (under reducing and non-reducing conditions) and isoelectric focusing varied considerably among BJC samples from the five snakes. The amino-terminal sequences (residues 1-34) of individual BJC samples were compared with the sequence deduced from isolated cDNAs encoding alpha and beta chains of BJC. A high degree of homology was detected, although some residues differed from one sample to other. Altogether, data confirmed the heterogeneity found for BJC purified from individual snakes. Thus, the results indicate that: (1) individual specimens of Bothrops jararaca have different patterns of BJC isoform expression; and (2) it seems that genetic factors, at least in part, determine the variability found in BJC production.
从巴西矛头蝮蛇毒液中分离出的一种强效特异性凝血酶抑制剂——矛头蝮蛇凝血酶抑制剂(BJC),在成年蛇采集的个体毒液中发现了不止一种同工型。先前已发现蛇毒成分的变化与年龄、性别、地理来源、一年中的季节和饮食等因素有关。为了获得有关BJC同工型个体表达模式的更多信息,我们分析了同时从同一地理区域的成年雌蛇采集的5份巴西矛头蝮蛇个体毒液。正如预期的那样,粗毒液在SDS-PAGE上显示出相似的迁移模式。使用包括亲和色谱步骤(PPACK-凝血酶琼脂糖)的方法纯化BJC。注意到从个体毒液样品中获得的BJC量略有变化。在来自5条蛇的BJC样品中,凝血酶诱导的血小板聚集抑制以及在SDS-PAGE上的迁移模式(在还原和非还原条件下)和等电聚焦有很大差异。将个体BJC样品的氨基末端序列(第1-34位残基)与从编码BJCα和β链的分离cDNA推导的序列进行比较。检测到高度同源性,尽管一些残基在不同样品之间有所不同。总之,数据证实了从个体蛇纯化的BJC存在异质性。因此,结果表明:(1)巴西矛头蝮蛇的个体标本具有不同的BJC同工型表达模式;(2)似乎遗传因素至少部分决定了BJC产生中发现的变异性。