Acta biomedica scientifica Acta biomedica scientifica Acta biomedica scientifica 2541-9420 2587-9596 14521 10.12737/23416 Микробиология и вирусология Microbiology and Virology Микробиология и вирусология detection of membrane-bound proteases of FRANCISEllA TUlARENSIS выявление мембраносвязанных протеаз FRANCISEllA TUlARENSIS Мазепа Андрей Владимирович Mazepa Andrey Владимирович Марков Евгений Юрьевич Markov Evgeniy Юрьевич Козлов Станислав Николаевич Kozlov Stanislav Николаевич Николаев Валерий Борисович Nikolaev Valeriy Борисович Корнева Александра Владимировна Korneva Aleksandra Vladimirovna adm@chumin.irkutsk.ru Попова Юлия Олеговна Popova Yuliya Олеговна Иркутский ордена Трудового Красного Знамени научно-исследовательский противочумный институт Иркутск Россия Irkutsk Antiplague Research Institute Irkutsk Russian Federation 11 12 2016 11 12 2016 1 5 155 159 https://zh-szf.ru/en/nauka/article/14521/view

Проведено исследование наличия протеолитической активности и состав протеаз в поверхностных структурах клеток шести штаммов Francisella tularensis разных подвидов. Препараты мембранной фракции возбудителя туляремии получали путем обработки микробной массы мочевиной. Субстратный электрофорез выявил три общих для всех штаммов полипептида, обладающих протеолитической активностью. У препаратов F.tularensis subsp. nearctica и F.tularensis subsp. novicida обнаружены дополнительные полосы протеаз.

Sterile preparations of membrane fractions were prepared by processing of live Francisella tularensis cells of different subspecies with 4.5M urea solution and differential centrifugation. For the first time, proteolytic activity was detected and studied by tests of radial enzyme diffusion and substrate polyacrylamide gel electrophoresis using gelatin as a substrate. Spectrum of gelatinases in the resulted preparation were detected. Quantitative inter-strain differences in the protease activities and their qualitative composition in membrane preparations of various virulent F.tularensis strains was analyzed. Avirulent F.tularensis 21/400 subsp. holarctica (I-214) strain demonstrated the greatest gelatinase activity in enzyme diffusion method and the lowest hydrolytic activity was seen in F.tularensis B-399 A-Cole subsp. tularensis (I-386) and F.tularensis Utah 112 subsp. novicida (I-384), other preparations showed intermediate activity. Enzyme electrophoresis in the protease spectra determined the presence of proteins with proteases activity 50–100kDa, and in the spectrum preparations of F. tularensis I-386 and I-384 were detected additional bands of proteases.

 Francisella tularensis субклеточные фракции наружные мембраны ферменты протеазы протеолитическая активность Francisella tularensis subcellular fractions outer membrane enzyme protease proteolytic activity

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