Makevnina L G, Golubenko Z, Akhunov A, Paskhina T S
Biokhimiia. 1990 Aug;55(8):1481-90.
The nature of the bradykinin (BK)-hydrolyzing (kininase) activity of peptidhydrolase isolated from spider (Latr. tredecimguttatus) venom has been studied. It was found that the BKase activity of the enzyme is fully inhibited by organic mercurials (10(-5)-10(-6) M) as well as by 5,5'-dithiobis(2-nitrobenzoic acid) (10(-7) M); the latter blocks three SH-groups within the enzyme molecule. Serine and metalloproteinase inhibitors have no effect on the kininase activity. Thin-layer chromatography on silicagel revealed that the highly purified enzyme hydrolyzes the -Pro7-Phe8- bond of BK liberating the C-terminal dipeptide, HPhe-ArgOH. Besides, the kininase splits off the C-terminal tripeptide from angiotensin I by hydrolyzing its -Pro7-Phe8-bond. The enzyme does not exhibit any exopeptidase activity with free and N-substituted tri- and pentapeptides. The data obtained suggest that the Latr. tredecimguttatus kininase can be related to thiol endopeptidases hydrolyzing the peptide bonds formed by proline carboxyl.
对从蜘蛛(红斑寇蛛)毒液中分离出的肽水解酶的缓激肽(BK)水解(激肽酶)活性的性质进行了研究。发现该酶的BK酶活性完全被有机汞化合物(10^(-5)-10^(-6) M)以及5,5'-二硫代双(2-硝基苯甲酸)(10^(-7) M)抑制;后者会封闭酶分子内的三个巯基。丝氨酸和金属蛋白酶抑制剂对激肽酶活性没有影响。硅胶薄层层析显示,高度纯化的酶水解BK的-Pro7-Phe8-键,释放出C末端二肽HPhe-ArgOH。此外,激肽酶通过水解其-Pro7-Phe8-键从血管紧张素I上裂解下C末端三肽。该酶对游离的和N-取代的三肽及五肽不表现出任何外肽酶活性。所获得的数据表明,红斑寇蛛激肽酶可能与水解由脯氨酸羧基形成的肽键的巯基内肽酶有关。