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Biomedical subjects

A A Baev

Publications and source records attributed to A A Baev.

At least 19 recordsLinked to original sources
PubMed1990

[Structure of the Pseudomonas aeruginosa recA gene].

The nucleotide sequence of the 1206 bp fragment of Pseudomonas aeruginosa DNA coding for the recA gene has been determined. This structure was shown to contain an open reading frame corresponding to a protein with m.w. 36808 D highly homologous (70%) to the Escherichia coli recA protein. Homology on the DNA level is significantly lower (57%) due to the high G/C content characteristics of Pseudomonas DNA. Making use of S1 nuclease and reverse transcriptase it was shown that in P. aeruginosa and E. coli cells recAPA gene transcription starts from A or T unit. Unlike "-35" region, "-10" region is homologous to the consensus E. coli promoter sequence. Comparison of primary structures of the recAPA and recAEC proteins demonstrates that the recAPA protein is by 7 amino acid residues shorter and differs from recAEC at 108 positions. Homology is the lowest in the C-terminal part. Basing on the analysis of hybrid recAPA proteins with a modified C-terminal part, it may be suggested that C-terminus is nonessential for main activities of the recA protein.

Amino Acid Sequence↗
PubMed1979

[Isolation and properties of DNA-cytosine-methylase I from Escherichia coli MRE 600].

DNA-cytosine-methylase I was isolated and purified to homogeneity. The yield made up to about 30% of total activity. The enzyme molecular weight as determined by centrifugation in a sucrose gradient, by gel filtration and by electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate was found to be 45,000. The Michaelis constant was 1,8 . 10(-6) M for SAM and 2 . 10(-4) M for DNA. DNA-cytosine-methylase I modifies phage lambda DNA in 60 sites. This modification does not protect DNA from the effects of restriction endonucleases HpaII and BsuRI. The enzyme methylates DNA in the nucleotide sequence: 5'...Pur-MC-C-G-G-Pyr...3'.

DNA (Cytosine-5-)-Methyltransferases↗
PubMed1979

[Isolation and properties of DNA-cytosine-methyltransferase EcoRII and E. coli K12].

The methods of isolation and partial purification of two DNA-cytosine-methylases (DC-methylases) EcoRII and E. coli K12 are described. After chromatography on phosphocellulose the enzymes were purified 100-fold, the yield being 30%. Further purification of the enzymes was performed by sedimentation in a sucrose concentration gradient. Both enzymes have native molecular weights of 50,000; DC-methylase from E. coli K12 may simultaneously occur in the forms with molecular weights of 70,000, 90,000 and 110,000. Both DC-methylases modify identical nucleotide sequences of DNA, have equal numbers (90) of methylation sites in phage lambda DNA and provide in vitro a complete protection of phage lambda DNA against restriction endonuclease EcoRII. DC-methylases E. Coli K12 and EcoRII differ in their chromatographic behaviour on phosphocellulose and capacity to form compexes with the cell DNA-adenine-methylase.

Coliphages↗
PubMed1979

[New series of thermostable phage T5 mutants carrying deletions in the region of the tRNA genes].

Six mutants of phage T5 have been selected by temperature inactivation in the presence of chelate-forming agents. Hybridization of DNA of the mutants with 4S RNA of phage T5+ has shown that all mutants have the deletions affecting tRNA genes. The size of the deletions and their location about the sites of DNA clevage with endonucleases EcoR1, Hind III, PstI and the nicks have been determined.

Chromosome Deletion↗
PubMed1978

[Inhibitors of exonuclease A5].

Over 30 compounds resembling to or being structural elements of the minimal substrate of exonuclease A5 were tested for their ability to inhibit the reaction catalyzed by this nuclease. The compounds containing less than two phosphate groups were shown to possess a low inhibitory activity, if any. CDP, double-stranded DNA and nucleoside-3',5'-diphosphates (pNp) proved to be effective exonuclease A5 inhibitors. Pyrophosphate stimulated the reaction in the case of low molecular weight substrates only. For the inhibitory activity of pNp to occur, the intactness of the nucleoside moiety as a whole was shown to be necessary, the activity level depending on the structure of both the base and the sugar components. A competitive mechanism of the inhibitory action was demonstrated for pTp, pdCp, pdGp, pdAp and pUp and the Ki values were determined. The affinity for the inhibitor decreased in the following order: pdCp greater than or equal to pTp greater than pdGp greater than pdAp. Ki for pdCp and pTp were found to be approximately 3.10(-6) M. The investigation of the inhibition mechanism as well the determination of Ki were accomplished with the help of homogenous low molecular weight substrates--ApApA and the phosphoamide MeOPheNH(pdA)2. These were chosen after kinetic parameters determination of the hydrolysis of 22 exonuclease A5 substrates, predominatly of the RpNpN type. On the basis of data obtained the specificity of exonuclease A5 is also discussed. Possible usefulness of immobilized competitive inhibitors of the pNp type not only for single nuclease isolation but for the separation of a mixture of different nucleases is considered. This possibility is based on the almost universal inhibitory effect of pNp on different nucleases and at the same time on their different affinity for the enzymes. In particular, this approach might be useful for the elimination of exonucleases and some other nucleolytic enzymes from the preparations of endonucleases-restrictases.

Cytidine Diphosphate↗