Analysis of ligand-receptor binding by the difference method

A new method has been proposed for analysis of experimental data on ligand-receptor binding at equilibrium. This method makes it possible to detect heterogeneity of a receptor system in cases where the contribution of the high-affinity site to total binding is rather small and the problem of graphic discrimination of a model cannot be solved unambiguously by other methods. The difference method permits us to exclude experiments on measuring nonspecific binding. A computer program for analysis of ligand-receptor binding has been worked out in which the difference method and traditional methods of binding isotherm analysis are realized. Numerical modeling has shown that the best strategy in experimental data processing is the treatment of total binding isotherms by both the difference method and regression analysis, including the nonspecific binding constant as one of the regression parameters.     

Complete assignment of signals in 1D and 2D H-NMR spectra of a 17-member oligonucleotide, a model symmetrical analog of lambda operators

A synthetic analog of lambda phage operators, a symmetric duplex of oligonucleotides, was studied by 1H NMR at 400 MHz. Signals in the spectrum of imino protons of the duplex in H2O were assigned based on the results of NOE experiments and temperature dependences. Resonance assignment of the non-exchangeable protons of bases and deoxyribose was performed by analysing the NOESY spectrum obtained in the single experiment. The results indicate that the major part of the duplex has a conformation similar to the B-form of DNA, and the region of the central non-complementary base pair exhibits deviations from the regular structure.     

1H-NMR study of the OR1 operon in bacteriophage lambda. I. Assignment of signals from imino protons and adenine C2 protons

An oligodeoxyribonucleotide composed of 17 residues, d(TATCACCGCCAGAGGTA), and a complementary chain were synthesized. Their duplex was identical with the operator OR1, the binding site for bacteriophage lambda cro and c1 repressors. The 1H NMR spectra (500 MHz) of the duplex imino and aromatic protons were studied at 10, 20 and 25 degrees C. Signals from the imino protons of complementary base pairs and from the C2 protons of adenine (with the exception of the duplex terminal nucleotides) were assigned using the NOE technique and the known characteristics of short DNA fragment melting. No signals from the imino protons of the terminal base pairs were detected even at 10 degrees C due to fraying which increased as the temperature was raised. The assignment of signals can be used to identify centers of interaction between the operator OR1 and repressors, as well as to study possible local changes in DNA geometry.     

Spatial structure of cro-repressor in a solution. II. Effect of ionized groups

The technique of 1H NMR spectroscopy and absorption UV spectroscopy were used to study the ionization of the tyrosine phenol cycles and the effect of ionizable groups on the chemical shifts of signals from protons in the side chains of several amino acid residues. The microenvironment of these residues was established by analysing the titration curves. The mutual orientation of two functionally important adjacent alpha-helical protein regions was determined in solution. The signals from methionine residues belonging to different regions of the secondary structure were assigned in the NMR spectrum. The results indicate that the spatial structure of the repressor is similar in solution an in the crystal. They confirm the model proposed for the cro repressor interaction with DNA and based on the data of X-ray diffraction analysis.     

An embryonic enantiornithine bird and associated eggs from the cretaceous of Mongolia

Enantiornithes is the most speciose clade of Cretaceous birds, but many taxa are known from isolated postcranial skeletons. Two embryonic enantiornithine bird skeletons of Gobipipus reshetovi gen. et sp. nov. from the Upper Cretaceous (Campanian) Barun Goyot Formation of the Gobi Desert in Mongolia provide new insights into the anatomy, radiation, and mode of development of early avialans. In recent times, both enantiornithine and ornithuromorph birds are known from the Barun Goyot Formation as well as from the Djadokhta and Nemegt Formations. The 80-million-year-old Gobipipus skeletons encased within eggshells shows several features characteristic of enantiornithine birds. The wing skeleton and shoulder girdle show morphological features indicating that Gobipipus achieved sophisticated powered flight. Gobipipus reshetovi gen. et sp. nov. is quite distinct from the sympatric enantiornithine species Gobipteryx minuta from the same strata in many anatomical features. Phylogenetic analysis of 26 avialan ingroup taxa based on distribution of 202 characters indicate that Gobipipus is a basal member of enantiornithine birds along with Confuciusornis and shares more characters with ornithuromorphs than previously recognized. The embryonic nature of Gobipipus specimens sheds new light on the developmental history of enantiornithine birds. The well-ossified bones of the fore- and hind limbs, and fusion of many skeletal elements indicate a precocial mode of development in Gobipipus. Apparently Gobipipus hatchlings could walk away from the ground nests as soon as they emerged from their eggs. The asymmetry of egg poles are unique features of Gobipipus eggs (oogenus Gobioolithus) among Cretaceous avialans. The microstructure of the shell in Gobioolithus eggs with the embryos of Gobipipus is typical avian (of ornithoid basic type) and less ratite-like in morphology of the spongy layer than is that in the other possible egg-remains of enantiornitine birds (oofamily Laevisoolithidae).     

Conjugates of 2',3'-didehydro-3'-deoxythymidine with thymogen. Synthesis and anti-HIV activity

An effective synthesis of thymogen was developed. Conjugates of 2',3'-didehydro-3'-deoxythymidine (nucleoside d4T) with thymogen were prepared in which the nucleoside hydroxyl group was linked to the thymogen carboxyl group of either tryprophan or glutamic acid residues. It was shown that the anti-HIV activity of the d4T-thymogene conjugate with the tryptophan linkage was comparable to that of d4T, whereas its cytotoxicity was nil. The d4T-tryptophan conjugate also displayed high anti-HIV activity.     

New species of sap beetles (Coleoptera: Nitidulidae: Nitidulini) from the Baltic and Bitterfeld ambers

A new genus, Microsoronia, gen. nov., and new species of this genus, M. hoffeinsorum, sp. nov. from the Bitterfeld amber and M. kerneggeri sp. nov., M. nigerrima sp. nov., and M. interfax, sp. nov. from the Baltic amber, are described. The earliest known member of the genus Phenolia, P. (Lasiodites) angustitibialis, sp. nov., is described from the Baltic amber. The systematic position of these two genera, their possible evolution, as well as the possible ecology and bionomics of their members are discussed. It is shown that “Phenolia” incapax Scudder, 1890 should be included in the family Peltidae, rather than Nitidulidae.     

Novel peroxisomal protease Tysnd1 processes PTS1- and PTS2-containing enzymes involved in beta-oxidation of fatty acids

Peroxisomes play an important role in beta-oxidation of fatty acids. All peroxisomal matrix proteins are synthesized in the cytosol and post-translationally sorted to the organelle. Two distinct peroxisomal signal targeting sequences (PTSs), the C-terminal PTS1 and the N-terminal PTS2, have been defined. Import of precursor PTS2 proteins into the peroxisomes is accompanied by a proteolytic removal of the N-terminal targeting sequence. Although the PTS1 signal is preserved upon translocation, many PTS1 proteins undergo a highly selective and limited cleavage. Here, we demonstrate that Tysnd1, a previously uncharacterized protein, is responsible both for the removal of the leader peptide from PTS2 proteins and for the specific processing of PTS1 proteins. All of the identified Tysnd1 substrates catalyze peroxisomal beta-oxidation. Tysnd1 itself undergoes processing through the removal of the presumably inhibitory N-terminal fragment. Tysnd1 expression is induced by the proliferator-activated receptor alpha agonist bezafibrate, along with the increase in its substrates. A model is proposed where the Tysnd1-mediated processing of the peroxisomal enzymes promotes their assembly into a supramolecular complex to enhance the rate of beta-oxidation.