Рекомбинантные пептиды Ce1 и Ce4 из яда скорпиона Centruroides elegans и их взаимодействие с гибридными каналами KcsA-K _V 1.X (x = 1, 3, 6)

Разработана методика получения рекомбинантных функционально-активных пептидов Ce1 и Ce4 из яда скорпиона Centruroides elegans в системе экспрессии Escherichia coli. Выходы Ce1 и Ce4 составили соответственно 6,5 и 12 мг с литра культуры (плотность клеток около 5 оптических единиц). Свойства полученных пептидов исследованы с применением биоинженерных систем на основе гибридных каналов KcsA-K_v1.x (х = 1, 3, 6), содержащих сайты связывания блокаторов соответствующих эукариотических калиевых каналов семейства K_v1. Показано, что рекомбинантные Ce1 и Ce4 вплоть до микромолярных концентраций не проявляют сродства к сайтам связывания каналов K_v1.1 и K_v1.6 и, подобно природным пептидам, селективно взаимодействуют с сайтом связывания канала K_v1.3: кажущиеся константы диссоциации комплексов KcsA-K_v1.3 с рекомбинантными Ce1 и Ce4 равны, соответственно, 50±10 и 200±30 нМ (среднее ± стандартная ошибка среднего).

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