The inhibition of pancreatic ribonuclease by anionic polymers

Fellig, J.; Wiley, Christine E.

doi:10.1016/0003-9861(59)90496-5

Cited by 82 publications

(18 citation statements)

References 11 publications

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“…As the dose of the polymer was increased, the stimulation first rose and then declined sharply to virtual extinction. The initial rise may be explained by inhibition of nuclease activity by dextran sulphate [24], an effect which has been observed directly in this system [14a]. A similar effect was apparent in the control nonpreincubated system, although the magnitude of the increase was smaller.…”

Section: Action Of Inhibitorssupporting

confidence: 57%

Mammalian Cell‐Free Protein Synthesis Directed by Viral Ribonucleic Acid

Mathews

Körner

1970

European Journal of Biochemistry

250

100

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1.A cell-free system from mouse Krebs I1 ascites cells is described which responds by increased amino acid incorporation into protein on addition of encephalomyocarditis virus (EMC-RNA). RNA from other sources does not produce this response.2 . The stimulation by EMC-RNA occurs over a narrow range of Mg2f concentrations and is maximal a t 5mM, which is optimal for the endogenous incorporation and lower than that required in the presence of poly(U).3. The EMC-RNA-directed product is distinguished from the endogenous products by its size and composition.4. After an initial lag of 4 min, during which the nascent chains are too short to be acidinsoluble, EMC-RNA-directed protein synthesis is active for more than 1 h.5. EMC-RNA-directed synthesis is particularly sensitive to the inhibitors cycloheximide and dextran sulphate. Using the latter, it has been shown that the initiation of protein synthesis is completed during the first 15 min of incubation.6. Only 20-3001, of the product of the cell-free system, whether endogenous or EMC-RNAdirected, is released from the ribosomes. Despite the size of the viral RNA, it did not appear to promote the formation of exceptionally large polysomes.7. It is concluded that the EMC-RNA is acting as a message for viral proteins in the ascites cell-free system. The demonstration that bacteriophage RNA can act as a messenger RNA in cell-free preparations from Escherichia coli [l] has greatly facilitated the study of several aspects of protein biospthesis in bacteria. We wished to extend this approach to mammalian systems using a homologous viral RNA. It is known that the RNA extracted from encephalomyocarditis virus (EMC) [2,3] and from several other mammalian viruses [4-61 is capable of stimulating amino acid incorporation when added to extracts of certain mammalian or avian cells and we have used the EMClKrebs I1 ascites cell system in this work. We report here the development of a highly sensitive cell-free preparation from mouse Krebs I1 ascites cells and describe some of the characteristics of the stimulation of protein synthesis elicited by encephalomyocarditis virus RNA (EMC-RNA). The fidelity of this system has recently been established by the demonstration that the product formed Unusual Abbreviations. EMC, encephalomyocarditis; Guo-5'-P2-CH,-P, 5'-guanylyl-methylenediphosphate; poly-(U), polyuridylic acid; S-30, the 30000 x g supernatant.Enzymes. Creatine phosphokinase (EC 2.7.3.2); pancreatic ribonuclease (EC 2.7.7.16).under the direction of EMC-RNA corresponds to material present in EMC-infected cells [7].

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Section: Action Of Inhibitorssupporting

confidence: 57%

Mammalian Cell‐Free Protein Synthesis Directed by Viral Ribonucleic Acid

Mathews

Körner

1970

European Journal of Biochemistry

250

100

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“…We were surprised to learn that 40 years ago, even poly(vinylsulfonic acid) had been tested as an inhibitor of RNase A. Those data suggested that poly(vinylsulfonic acid) was a worse inhibitor of RNase A than other polyanions (33,34), or alternatively, that only long polymers (Ͼ9,000 g/mol) were good inhibitors of RNase A (35). We do not know the basis for the disparity with our data.…”

Section: Discussionmentioning

confidence: 99%

Potent Inhibition of Ribonuclease A by Oligo(vinylsulfonic Acid)

Smith

Soellner

Raines

2003

Journal of Biological Chemistry

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Ribonuclease A (RNase A) can make multiple contacts with an RNA substrate. In particular, the enzymatic active site and adjacent subsites bind sequential phosphoryl groups in the RNA backbone through Coulombic interactions. Here, oligomers of vinylsulfonic acid (OVS) are shown to be potent inhibitors of RNase A that exploit these interactions. Inhibition is competitive with substrate and has K i ‫؍‬ 11 pM in assays at low salt concentration. The effect of salt concentration on inhibition indicates that nearly eight favorable Coulombic interactions occur in the RNase A⅐OVS complex. The phosphonic acid and sulfuric acid analogs of OVS are also potent inhibitors although slightly less effective. OVS is also shown to be a contaminant of MES and other buffers that contain sulfonylethyl groups. Oligomers greater than nine units in length can be isolated from commercial MES buffer. Inhibition by contaminating OVS is responsible for the apparent decrease in catalytic activity that has been observed in assays of RNase A at low salt concentration. Thus, OVS is both a useful inhibitor of RNase A and a potential bane to chemists and biochemists who use ethanesulfonic acid buffers.

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“…In these interactions, factors such as the conformation of the compounds, the size of the micelles in solution, etc., probably predominate so that the role played by the sulfate content becomes insignificant. Sulfanilylbenzamide, which is similar to the derivatives prepared here, has been shown to be effective against the bacillary dysentery organism (1) and against pneumococcus in mice (2).…”

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confidence: 81%