Mechanistic insights into RNase L through use of an MDMX-derived multi-functional protein domain

Garvie, Colin W.; Vasanthavada, Keshav; Xiang, Qing

doi:10.1016/j.bbapap.2013.04.010

Cited by 7 publications

(8 citation statements)

References 18 publications

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“…Some ribonucleases, such as RNase L and BS RNase, were shown to be natural dimers [29,30], with dimer ization necessary for their functioning. Thus, RNase L, a component of the cell response to viral infection, is activated by dimerization [29,31].…”

Section: Resultsmentioning

confidence: 99%

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Preparations of Bacillus pumilus secreted RNase: One enzyme or two?

et al. 2015

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Immunochemical analysis of the following purified preparations of Bacillus pumilus RNase (binase) was carried out: industrially manufactured enzyme (Institute of Organic Synthesis, Riga, Latvia) and the enzymes isolated from the culture liquid of the native B. pumilus producer and from the Escherichia coli BL21 recombinant strain bearing the pGEMGX1/ent/Bi plasmid. Electrophoresis of all three samples of purified binase revealed two protein fractions with ribonuclease activity possessing molecular masses of ~12 and 25 kDa. The possible presence of binase II, a second secreted RNase, was ruled out. Both high and low molecular mass proteins interacted with binase specific antibodies in the immunoblotting reaction, which indicated their antigenic identity. The difference in molecular mass between these proteins indicated the pos sible presence of two forms of binase in solution, a monomer and a dimer.

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Section: Resultsmentioning

confidence: 99%

“…Thus, RNase L, a component of the cell response to viral infection, is activated by dimerization [29,31]. BS RNase, which is not susceptible to the cytosolic RNase inhibitor [30], forms two types of dimers, which occur in a 2 : 1 ratio [32].…”

Section: Resultsmentioning

confidence: 99%

Preparations of Bacillus pumilus secreted RNase: One enzyme or two?

et al. 2015

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Section: результаты и обсуждениеunclassified

“…Показано, что некоторые рибонуклеазы, такие как РНКаза L и BS РНКаза, являются природны ми димерами [29,30], и димеризация необходима для выполнения их функций. Так, РНКаза Lкомпонент ответной реакции клеток на вирусную инфекцию, активируется с помощью димериза ции [29,31]. BS РНКаза, не подверженная дей ствию цитозольного ингибитора РНКаз [30], об разует два типа димеров, которые встречаются в соотношении 2 : 1 [32].…”

Section: результаты и обсуждениеunclassified

Препараты секретируемой РНКазыBacillus pumilus: один фермент или два?

et al. 2015

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Казанский (Приволжский) федеральный университет, Институт фундаментальной медицины и биологии, кафедра микробиологии Поступила в редакцию 13.08.2014 г. Проведен иммунохимический анализ очищенных препаратов РНКазы Вacillus pumilus (биназы), представленных ферментом заводского производства (Институт органического синтеза, Рига, Лат вия), ферментами, выделенными из культуральной жидкости нативного штамма продуцента В. pumilus и рекомбинантного штамма Escherichia coli BL21, несущего плазмиду pGEMGX1/ent/Bi. Установлено, что при электрофорезе всех трех образцов очищенной биназы обнаруживаются две белковые фракции, обладающие рибонуклеазной активностью. Определена молекулярная масса фракций-соответственно, около 12 и 25 кДа. Исключена возможность присутствия в препаратах второй секретируемой РНКазы В. pumilus-биназы II. Как низкомолекулярный, так и высокомоле кулярный белки взаимодействовали со специфическими антителами к биназе в реакции имму ноблоттинга, что указывает на их антигенную идентичность. Различие в молекулярной массе ис следуемых белков свидетельствует о том, что биназа в растворе может быть представлена двумя формами-мономером и димером.

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“…Although only one native dimeric RNase, the bull semen RNase (BS-RNase), has been identified to date [16] , RNase A can also form dimers under certain conditions [17] . Dimerisation is necessary for efficient RNA hydrolysis by RNase T from E. coli [18] , for the specific decay of viral RNA by RNase L participating in innate immunity against microbial pathogens [19] and by monocyte chemoattractant protein 1-induced protein 1 (MCPIP1) [20] , for the evasion of ribonuclease inhibitor RI by BS-RNase [21] and artificial dimers of RNase A [22] . We assumed that the ability of RNases to form dimers is a natural property that is more widespread than previously believed.…”

Section: Introductionmentioning

confidence: 99%

New Insight into Secreted Ribonuclease Structure: Binase Is a Natural Dimer

et al. 2014

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The biological effects of ribonucleases (RNases), such as the control of the blood vessels growth, the toxicity towards tumour cells and antiviral activity, require a detailed explanation. One of the most intriguing properties of RNases which can contribute to their biological effects is the ability to form dimers, which facilitates efficient RNA hydrolysis and the evasion of ribonuclease inhibitor. Dimeric forms of microbial RNase binase secreted by Bacillus pumilus (former B. intermedius) have only been found in crystals to date. Our study is the first report directly confirming the existence of binase dimers in solution and under natural conditions of enzyme biosynthesis and secretion by bacilli. Using different variants of gel electrophoresis, immunoblotting, size-exclusion chromatography and mass-spectrometry, we revealed that binase is a stable natural dimer with high catalytic activity.

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Mechanistic insights into RNase L through use of an MDMX-derived multi-functional protein domain

Cited by 7 publications

References 18 publications

Preparations of Bacillus pumilus secreted RNase: One enzyme or two?

Preparations of Bacillus pumilus secreted RNase: One enzyme or two?

Препараты секретируемой РНКазыBacillus pumilus: один фермент или два?

New Insight into Secreted Ribonuclease Structure: Binase Is a Natural Dimer

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