共查询到20条相似文献,搜索用时 15 毫秒
1.
A new endoribonuclease activity, RNase F, was partially purified from cells. This activity can cleave a precursor RNA molecule (of Species 1), isolated from T4 infected cells, in a specific site. This activity is different from the other three know processing endoribonucleases of RNase III, RNase E and RNase P. 相似文献
2.
Jerard Hurwitz Sue Wickner Michel Wright 《Biochemical and biophysical research communications》1973,51(2):257-267
Purified RNA polymerase, DNA polymerase III and unwinding protein of catalyze limited rifampicin sensitive fd or ØX 174 DNA-dependent DNA synthesis. A protein has been partially purified from which stimulates rifampicin sensitive dXMP incorporation in this system 20 to 30 fold. This protein also stimulates DNA synthesis catalyzed by DNA polymerases I and II; the stimulation occurs in reactions primed with natural and synthetic DNAs as well as RNA-DNA hybrids. The protein is not a product of the known dna genes. In contrast to the above system of purified enzymes, rifampicin sensitive dXMP incorporation in crude extracts of is specifically dependent on fd but not ØX 174 DNA. An additional factor has been isolated from extracts of which restores specificity to the purified rifampicin sensitive system by preventing ØX 174 DNA from serving as a template. 相似文献
3.
Are all nucleotidyl transferases metalloenzymes? 总被引:5,自引:0,他引:5
P Valenzuela R W Morris A Faras W Levinson W J Rutter 《Biochemical and biophysical research communications》1973,53(3):1036-1041
4.
C R Geren L M Geren K E Ebner 《Biochemical and biophysical research communications》1975,66(1):139-143
A thermostable protein that strongly inhibits the soluble D-alanine carboxypeptidase was isolated from a cell-free extract of . The inhibitor was purified 140-fold by heat treatment, selective precipitation at pH 4.5, ion exchange chromatography on DEAE-cellulose and gel chromatography on Sephadex G-100. Inhibition of soluble D-alanine carboxypeptidase by this inhibitor is reversed by cations such as Mg++ or Na+ and abolished by digestion of the inhibitor with proteolytic enzymes. The inhibitor does not affect either the particulate D-alanine carboxypeptidase of or the growth of the bacteria. 相似文献
5.
David R. McNaughton Glen R. Klassen Herb B. LéJohn 《Biochemical and biophysical research communications》1975,66(2):468-474
Three phosphorylated guanosine derivatives designated HS-1, HS-2 and HS-3 synthesised during active protein synthesis in the water-mould, sp (1969) were shown to regulate the enzymatic activities of nucleoplasmic and nucleolar DNA-dependent RNA polymerases (RNAP-I, II and III) from both and another unrelated water-mould, . These HS compounds were without effect on DNA-dependent RNA polymerase holoenzyme. The most potent of the three compounds was HS-3 which inhibited the activity of all enzymes completely at 100 μg/ml. HS-1, on the other hand, activated maximally at 1 to 10 μg/ml. HS-1 activation (3-fold) was restricted to enzyme III, and it had only partial inhibitory effects on enzymes I and II. The pattern of synthesis of HS-compounds throughout the 20-hour asexual growth cycle of the organism correlated with the detectable levels of the different RNA polymerases of . 相似文献
6.
F R Althaus S D Lawrence G L Sattler H C Pitot 《Biochemical and biophysical research communications》1980,95(3):1063-1070
The synthesis of proteins was examined, by two-dimensional O'Farrell gels, in mutant strains defective in all possible combinations of the RNA processing enzymes RNase III, RNase E and RNase P. We found that the synthesis of most proteins was unaffected; however, the synthesis of a significant number of proteins, 21 out of 80 tested, was drastically reduced in the strain defective in all three enzymes. It appears that the two enzymes RNase III and RNase E are responsible for most of these changes. 相似文献
7.
S Sakakibara F D Wicks R K Gholson 《Biochemical and biophysical research communications》1977,76(1):158-166
contains two proteins (A and B) which together convert dihydroxyacetone phosphate and aspartate to quinolinic acid, a precursor of NAD. Although mammalian liver homogenate does not catalyze this reaction it contains a protein which will replace the B protein of the system. The behavior of the liver protein on Sephadex G-75 suggests it is much smaller than the B protein. Liver B protein also appears to contain tightly bound FAD while FAD is easily removed from the B protein. The pH optimum for the hybrid system A protein-liver B protein is 9.0 while in the pure system the optimum is pH 8.0. The hybrid system is inhibited by NAD to the same extent as the pure system. 相似文献
8.
A strain of Escherichia coli lacking RNAase III and containing thermolabile RNAase E and RNAase P was labeled with 32Pi at a non-permissive temperature. RNA molecules were separated by two-dimensional polyacrylamide gel electrophoresis. Most of the small RNA species were isolated and analyzed for the presence of 5′ nucleoside triphosphates. In 16 of the 22 species analyzed a significant number of the individual molecules contained 5′ di or triphosphates. We conclude, therefore, that very little endonucleolytic RNA processing occurs in the absence of the three RNA processing enzymes RNAase III, RNAase E and RNAase P. 相似文献
9.
O. Bojan M. Bologa G. Niac N. Palibroda E. Vargha O. Bârzu 《Analytical biochemistry》1980,101(1):23-25
The application of a number of immobilized Procion dyes to the purification of inosine 5′-monophosphate dehydrogenase (EC 1.2.1.14) from Escherichia coli has been investigated. The enzyme is adsorbed to a number of these immobilized dyes and can be eluted by salt gradients with very substantial increases in specific activity. For example, adsorption of the enzyme from a crude cell-free extract of E. coli to immobilized Procion yellow MX-8G in the presence of 15% () ethylene glycol and subsequent elution with a salt gradient yields an enzyme preparation approximately 90% pure by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme is quantitatively recovered with an overall increase in specific activity of 14-fold compared to the enzyme in the cell-free extract. 相似文献
10.
J Sy N H Chua Y Ogawa F Lipmann 《Biochemical and biophysical research communications》1974,56(3):611-616
Ribosomes obtained from (ATCC 8185) were slightly active in synthesizing guanosine polyphosphates, which activity was greatly stimulated by addition of stringent factor. chloroplast ribosomes did not produce guanosine polyphosphates on incubation but responded with abundant synthesis to addition of the stringent factor from . In contrast, cytoplasmic ribosomes from the same organism did not respond. Interchange experiments between either subunit from chloroplasts with the counterparts showed good activity. When the small subunit of cytoplasmic ribosomes was combined with the large subunit of or of chloroplasts, a small but definite response was obtained. 相似文献
11.
S Nasu F D Wicks S Sakakibara R K Gholson 《Biochemical and biophysical research communications》1978,84(4):928-935
Two proteins (A and B) from are required for the synthesis of the NAD precursor quinolinate from aspartate and dihydroxyacetone phosphate. Mammalian liver contains a FAD linked protein which replaces B protein for quinolinate synthesis. D-aspartic acid but not L-aspartic acid is a substrate for quinolinic acid synthesis in a system composed of the B protein replacing activity of mammalian liver and A protein. In contrast the B protein- A protein quinolinate synthetase system requires L-aspartic acid as substrate. The previous report that L-aspartate was a substrate in the liver- system was due to contamination of commercially available [14C]L-aspartate with [14C]D-aspartate. These and other observations suggest that liver B protein is D-aspartate oxidase and B protein is L-aspartate oxidase. 相似文献
12.
R A Jacobson L F Mazzuckelli 《Biochemical and biophysical research communications》1973,54(3):867-873
Two DNA-dependent RNA polymerases have been isolated from , a bacterium with a distinct morphological life cycle. The two enzymes are different with respect to chromatographic and electrophoretic behavior, divalent cation requirement, and template activity with different DNA species. One appears to be similar in its properties and structure to polymerase, the other is different, but the nature of the difference is not yet clear. The possible relationship of the two enzymes in a regulatory role in the life cycle has yet to be investigated. 相似文献
13.
GMP synthetase was purified 180-fold from B and 18-fold from the derepressed purine auxotroph, B-96. The enzymes from both sources show the same preference for glutamine over ammonia as amino donor. Each is dimeric, consisting of subunits of molecular weight about 60,000. Thus the two are apparently identical. The similarities between GMP synthetase and xanthosine 5′-phosphate aminase of B-96 (N. Sakamoto, G.W. Hatfield, and H.S. Moyed, J. Biol. Chem. (1972) , 5880–5887) in respect to structure, state of derepression, and behavior during purification, lead us to the conclusion that the synthetase and the aminase are a single entity. We observe no loss or separation of glutamine-dependent activity upon purification of GMP synthetase and we suggest that such loss, reported by other workers, results artifactually by inactivation of an intrinsic glutamine-binding site. GMP synthetase appears not to contain a glutamine-binding subunit which is separable from the xanthosine 5′-phosphate-aminating component. 相似文献
14.
Philippe Régnier 《Biochemical and biophysical research communications》1981,99(3):844-854
The proteolytic activity of measured using 125I-labelled αS1 casein as substrate, is mainly localised in the outer membrane and is due to an intrinsic outer membrane protein which can be solubilized by deoxycholate. This enzyme exhibits maximum activity at pH 7,5 in Tris-HCl buffer, is resistant to thermal denaturation with a half-life of 28 min. at 90°C in deoxycholate-NaCl buffer and is inhibited by ethylene-diamine tetraacetate, high concentrations of p-aminobenzamidine, tosyl-L-lysine chloromethyl ketone, tosyl-L-phenylalaninechloromethyl ketone and by two inhibitors of the processing of the secreted protein precursors, procaine and phenehylalcohol. Whole cells do not exhibit proteolytic activity, nevertheless, some is unmasked when the outer membrane is permeabilized by Tris or ethylenediamine tetraacetate or when vesicles are sonicated. This suggests that the protease is on the inner side of the outer membrane. Because the protease is different from the soluble proteases described in , and especially from proteases I,II and III, it has been called protease IV. 相似文献
15.
A polyphosphate kinase has been purified over 100-fold from an extract of K-12. It requires both orthophosphate and a basic protein (histone or protamine) for maximum activity. Because its activity is stimulated by histone, polyphosphate kinase may easily lead to an error in the determination of protein kinase in the cell extract. Our data suggest that the stimulatory effect of orthophosphate on polyphosphate kinase may be important in the regulation of phosphate metabolism in the microorganism. 相似文献
16.
S Friedman 《Biochemical and biophysical research communications》1979,89(4):1328-1333
5-Azacytidine, when added to growing K12, causes a decrease in DNA methylation assayed . This decrease is greater when DNA is used as substrate than when calf thymus DNA is used. The decrease in activity is not due to the inhibition of protein synthesis caused by this drug, since neither chloramphenicol nor rifampin causes a decrease in enzyme activity. The effect is specific for the DNA(cytosine-5)methylase; the methylation of adenine is not affected. The concentration of drug that inhibits the DNA methylase by 50% is the same concentration that inhibits cell growth by 50%. 相似文献
17.
Chemically formylated Met-tRNAmMet and Met-tRNAfMet species from and yeast were tested for their capacity to serve as chain-initiators in a cell-free system from . In the presence of R 17 mRNA, initiation factors and ribosomes, all four Met-tRNAs could form functional initiation complexes as measured by ribosomal binding kinetics, fMet-puromycin formation and synthesis of a dipeptide fMet-Ala. Unformylated Met-tRNAfMet from displayed significantly less activity as a peptide chain-initiator than the formylated Met-tRNAmMet species from and yeast. Although the latter tRNAs were less effective initiators than the “physiological” initiator tRNAs, the data seem to indicate that a blocked α-amino group represents the major token of identification by which Met-tRNA is admitted to function in peptide chain initiation. 相似文献
18.
M C Graves C A Breitenberger L L Spremulli 《Archives of biochemistry and biophysics》1980,204(2):444-454
An improved method for the isolation of Euglena chloroplast ribosomes is described which presents a number of advantages over past procedures. First, ribosomes are prepared from whole cell extracts, thus bypassing the need to isolate intact chloroplasts and resulting in a 10-fold improvement in yield. Second, the inclusion of 40 mm Mg2+ in the preparation buffers, while stabilizing the chloroplast ribosomes, precipitates and, thereby, virtually eliminates the cytoplasmic 89 S ribosomes. Third, greater than 95% of the chloroplast ribosomes sediment at 68 S rather than as the damaged 53 S particle frequently generated in other preparation procedures. Fourth, even with a high-salt wash to remove endogenous factors, the chloroplast ribosomes still sediment at 68 S and are just as active in in vitro protein synthesis as are E. coli ribosomes. These ribosomes have been tested for activity with elongation factors from prokaryotes, eukaryotes, and the chloroplast itself, and the results have been compared to those obtained with E. coli and wheat germ ribosomes. The data may be summarized as follows: (a) Chloroplast ribosomes use E. coli and EF-G with the same efficiency as do E. coli ribosomes in protein synthesis, (b) E. coli and chloroplast ribosomes can use Euglena chloroplast EF-G to catalyze translocation, but wheat germ ribosomes cannot, (c) Wheat germ EF-1H and EF-2 are highly active in polymerization with wheat germ ribosomes, but ribosomes from neither E. coli nor the chloroplast are able to recognize these factors, (d) All three types of ribosomes accept Phe-tRNA from E. coli EF-Tu although to differing degrees. However, neither chloroplast nor E. coli ribosomes recognize wheat germ EF-1H for the binding of Phe-tRNA. 相似文献
19.
Two valine-sensitive acetohydroxy acid synthase activities were separable from K-12 cells by virtue of their different affinities for DEAE-cellulose eluted with a KC1 gradient. These activities appeared to be independent from a valine-resistant cryptic component expressed only in regulatory mutants. The properties of the first and second activity were coincident to those of extracts of and mutants, respectively. These data prove that the and gene products exist in the cell as physically distinct acetohydroxy acid synthase isoenzymes. 相似文献
20.
The absence of ribothymidine in specific eukaryotic transfer RNAs. I. Glycine and threonine tRNAs of wheat embryo 总被引:5,自引:0,他引:5
K Marcu R Mignery R Reszelbach B Roe M Sirover B Dudock 《Biochemical and biophysical research communications》1973,55(2):477-483
Ribothymidine, generally considered a universal nucleotide in tRNA, is completely absent in five specific wheat embryo tRNAs. These consist of two species of glycine tRNA and three species of threonine tRNA. These tRNAs, all extensively purified, are acceptable substrates for - ribothymidine forming-uracil methylase, which produces one mole of ribothymidine per mole of tRNA. These five tRNAs account for about 90% of the wheat embryo tRNAs which are substrates for this methylase. Nucleotide sequence analysis of one of these tRNAs, tRNAGlyI, confirmed both the complete absence of ribothymidine at position 23 from the 3′end, and the presence of uridine at that site instead. In addition, it is shown that methylation with uracil methylase quantitatively converts uridine at position 23 to ribothymidine, while no other uridine in the molecule is affected.Using uracil methylase as an assay we have detected this class of ribothymidine lacking tRNA, in each case consisting of a few specific species, in other higher organisms, such as wheat seedling, fetal calf liver and beef liver, in addition to wheat embryo. We could not detect this class of tRNA in or yeast tRNA. 相似文献