[Localization of binding sites of E. coli DNA-dependent RNA-polymerase with photosensitive template analogs].

The photoinduced covalent binding of E. coli RNA polymerase with decathymidylic templates containing 5-bromouracil residue has been carried out. Peptides from beta and beta' subunits of the core-enzyme, situated in the DNA-template binding site of the RNA polymerase active center have been localized.

[Preparation and characteristics of monoclonal antibodies against GTP-binding protein from the bovine retina].

Five monoclonal antibodies against the native GTP-binding protein (transducin) from bovine retina have been prepared. By immunoblotting and immunoenzymatic analysis of the isolated alpha- and gamma-subunits of transducin and the beta gamma-subunit complex it was determined that two monoclonal antibodies A3G7 and A3C10 recognize linear antigenic determinants on the alpha-subunit, two other, A3E4 and 3B3, bound specifically to the gamma-subunit, and monoclonal antibodies 1C3 interact only with native transducin. Both antibodies against the alpha-subunit inhibited transducin GTPase activity, whereas antibodies A3E4, 3B3 and 1C3 did not affect it.

[Primary structure of the gamma-subunit of the GTP-binding protein from the bovine retina].

The complete amino acid sequence of the gamma-subunit of GTP-binding protein from cattle retina has been established. The polypeptide chain consists of 69 amino acid residues and contains an unusual sequence Cys35-Cys36. The molecular mass of the gamma-subunit is 8008,7.

[Effective method of oligonucleotide-controlled mutagenesis of DNA fragments].

A procedure has been designed for changing specific nucleotides in a DNA sequence with efficiency. The method involves the use of the specially constructed cloning vectors pBRS1, pHS1, and pHS2. These plasmids are derivatives of pBR322 in which the EcoRI-HindIII region has been replaced by synthetic duplexes carrying SmaI, HpaI and XhoI sites, in addition to EcoRI and HindIII sites.

[Analysis of GTP-binding protein from the bovine retina. Complete amino acid sequence of the gamma-subunit].

The complete amino acid sequence of gamma-subunit of GTP-binding protein from bovine retina was determined. The polypeptide chain consists of 69 amino acid residues. It contains unusual sequence -Cys35-Cys36- with disulfide bridge.

[Expression in Escherichia coli cells of mutant human interferon alpha2].

cDNA library was obtained from mRNA isolated from human leukocytes induced by Newcastle disease virus. Clones containing cDNA for alpha 2-interferons were identified by colony hybridization with two synthetic hexadecanucleotides. One of the positive clones contained a NH2-terminal part of cDNA of human interferon identical to cDNA for IFN-alpha 2.

[Conformation NMR analysis of the spatial structure of Buthus eupeus insectotoxin I5A].

1H NMR spectroscopy has been used to collect data related to the spatial structure of insectotoxin I5A Buthus eupeus: pH-dependence of the chemical shifts, deuterium exchange rates of individual amide hydrogens, spin-spin coupling of the H-N-C alpha-H and H-C alpha-C beta-H protons, and nuclear Overhauser effect between distinct protons belonging to amino acid residues remote in the sequence. Molecular conformation in the regions from Asp9 to Cys19 (beta-turn 9-12 and right-hand alpha-helix 12-19) and from Asn23 to Asn34 (antiparallel beta-sheet with the beta-turn 27-30) directly follows from the observed parameters. Pseudoatomic approach of distance geometry algorithm was used to solve the overall folding of the molecule and propose the most probable set of disulfide bridges: Cys2-Cys19, Cys5-Cys31, Cys16-Cys26 and Cys20-Cys33.

[Visual rhodopsin. III. Complete amino acid sequence and topography in a membrane].

Tryptic hydrolysis of apomembranes, BNPS-skatole cleavage of carboxymethylated rhodopsin and thermolytic digestion of native membranes were carried out to obtain the peptides necessary for the polypeptide chain reconstruction. Gel-filtration on Bio-Gel P-30 in 80% formic acid, ion-exchange and reversed-phase high performance liquid chromatography were used for the peptide isolation. A comparison of rhodopsin hydrophobicity profile with the accessibility of the polypeptide chain in native photoreceptor membranes for proteases allowed to distinguish seven alpha-helical segments and propose a model for arrangement of the protein molecule in the membrane.

[Primary structure of the elongation factor G from Escherichia coli. IX. Structure of peptides generated by cyanogen bromide cleavage of the G-factor isolated on thiol-activated sepharose and of the products of the G-factor cleavage at Asp-Pro bonds. Complete primary structure].

The amino acid sequence of cysteine- and cystine-containing peptides resulting from cleavage of the G-factor by cyanogen bromide has been determined. For structure analysis cyanogen bromide peptides were further degradated using trypsin, chymotrypsin, thermolysin, staphylococcal glutamic protease, or limited acid hydrolysis. The products of the G-factor cleavage at Asp-Pro bonds were also studied.

[Comparative study of the biological properties of plasmid and natural human interferons].

A comparative study of biological properties of natural and plasmid human interferons was carried out. Natural and leukocyte interferons: alpha (induced by Newcastle disease virus) and gamma (induced by staphylococcal enterotoxin A) as well as natural fibroblastic beta interferon induced by poly(I) X poly(C) were studied in comparison with plasmid interferons alpha-F and alpha-F/D obtained from recombinant bacteria. Antigenic determinants of plasmid interferons alpha-F and alpha-F/D were found to be identical with those of natural and alpha-interferon of man and to differ from those of natural human alpha- and beta-interferons.