Kovacs K L, Seefeldt L C, Tigyi G, Doyle C M, Mortenson L E, Arp D J
Department of Biochemistry, University of Georgia, Athens 30602.
J Bacteriol. 1989 Jan;171(1):430-5. doi: 10.1128/jb.171.1.430-435.1989.
We examined the immunological cross-reactions of 11 different hydrogenase antigens with 9 different hydrogenase antibodies. Included were antibodies and antigens of both subunits of the hydrogenases of Bradyrhizobium japonicum and Thiocapsa roseopersicina. The results showed a strong relationship among the Ni-Fe dimeric hydrogenases. The two subunits of Ni-Fe dimeric hydrogenases appeared immunologically distinct: specific interactions occurred only when antibodies to the 60- and 30-kilodalton subunits reacted with the 60- and 30-kilodalton-subunit antigens. The interspecies cross-reactions suggested that at least one conserved protein region exists among the large subunits of these enzymes, whereas the small subunits are less conserved. Antibodies to the Fe-only bidirectional hydrogenase of Clostridium pasteurianum reacted with the Desulfovibrio vulgaris bidirectional hydrogenase. Surprisingly, antibodies to the clostridial uptake hydrogenase did not react with any of the Fe-only bidirectional hydrogenases but did react with several of the Ni-Fe dimeric hydrogenases. The two hydrogenases from C. pasteurianum were found to be quite different immunologically. The possible relationship of these findings to the structure and catalytic functions of hydrogenase are discussed.
我们检测了11种不同氢化酶抗原与9种不同氢化酶抗体之间的免疫交叉反应。其中包括日本慢生根瘤菌和玫瑰色硫球菌氢化酶两个亚基的抗体和抗原。结果显示镍铁二聚体氢化酶之间存在密切关系。镍铁二聚体氢化酶的两个亚基在免疫学上表现出明显差异：只有当针对60千道尔顿和30千道尔顿亚基的抗体与60千道尔顿和30千道尔顿亚基抗原发生反应时，才会出现特异性相互作用。种间交叉反应表明，这些酶的大亚基中至少存在一个保守的蛋白质区域，而小亚基的保守性较差。巴氏梭菌仅含铁双向氢化酶的抗体与普通脱硫弧菌双向氢化酶发生反应。令人惊讶的是，梭菌摄取氢化酶的抗体与任何一种仅含铁双向氢化酶均无反应，但与几种镍铁二聚体氢化酶发生反应。发现来自巴氏梭菌的两种氢化酶在免疫学上有很大差异。讨论了这些发现与氢化酶结构和催化功能之间的可能关系。
