Inactivation of microbial glutamin-(asparagin-)ase by azaserine and 6-diazo-5-oxo-L-norleucine

The effect of substrate analogues on glutamin-(asparagin-)ase from Pseudomonas aurantiaca-548 has been studied. The enzyme was demonstrated to be highly sensitive to the the action of 6-diazo-5-oxo-L-norleucine and azaserine. L-isomers of glutamine, aspartate, glutamate and several other substrate analogues with free alpha-amino groups protected the enzyme against the inhibitory DON effect. Thus, thorough preliminary selection of appropriate inhibitors, their dosage and treatment duration is needed for the recommendation of combined enzyme-inhibitor application in anti-tumour chemotherapy.     

Structural changes in contractile proteins of muscle fibers studied by polarization ultraviolet fluorescence microscopy. X. The effect of ATP, Ca2+, pH change and ionic strength of the washing solution on the structural state of thick filaments

By the method of polarized ultraviolet fluorescence microscopy, effects of ATP, Ca2+, changes in pH and ionic strength of washing solution of the structural state of thick filaments in both actin-free muscle fibers of rabbit and anisotropic discs (A-discs) of glycerinated fibers of crab were studied. The dependence of tryptophan fluorescence anisotropy of thin filaments upon physico-chemical parameters (compounds) of washing solution has been found. The structural state of thick filaments was suggested to be influenced by ATP, Ca2+, changes in pH and ionic strength of washing solution.     

A rapid immunofluorescent method of determining the antibiotic sensitivity of brucella

Optimal conditions for rapid assay of Brucella antibiotic sensitivity with the immunofluorescent method were developed. With this method high sensitivity of the main Brucella species to tetracycline, doxycycline and rifampicin was confirmed. It was found actually possible to use the immunofluorescent method for rapid assay of Brucella antibiotic sensitivity in practice.     

Structural changes in the contractile proteins of muscle fiber studied by polarization ultraviolet fluorescence microscopy. IX. The effect of the pH and ionic strength of the solution on the conformational restructurings of F-actin induced by the binding of heavy meromyosin

The dependence of F-actin conformational changes induced by the F-actin-HMM complex on pH and ionic strength was found by polarized ultraviolet fluorescence microscopy. It is discovered that pH affects sufficiently the cooperativity of F-actin structural changes, while the ionic strength affects their depth. The actomyosin complex was supposed to be at least in two structural states, differing in their orientation as well as in flexibility of F-actin monomers.     

Translocation of transposons Tn10 and Tn5 in the Yersinia pestis chromosome

The possibility of translocation of the transposons Tn5 and Tn10 into the genome of Yersinia pestis, with the subsequent mutagenic effect was demonstrated. We revealed transposon harbouring clones at frequency 10(-4) to 10(-2). Derivatives of P1cml clr100ts phage served as vectors. Insertion of Tn10 transposon induced mutations in ilv, ser, arg, pur, pro, leu, nic, tyr, gua genes. The number of the insertion sites on the chromosome obtained for Tn5 was the same, these being arg, ade, pyr, leu, gua, trp, his, pan, ilv. The majority of auxotrophs did not revert. Occasionally, revertants were observed at frequencies 10(-8) to 10(-6). Unlike Escherichia coli, reversion was not accompanied by the loss of transposons. The rearrangements induced by transposons, presumably, near the insertion site, as well as duplications of transposons followed by incorporation of copies into novel sites, led to the appearance of additional defective genes, which made it possible to select various types of polyauxotrophs. Based on reiteration of coinciding double and triple mutant markers, we proposed a linkage group of genes within a segment of Y. pestis chromosome: lys ... tyr - ser - arg - ilv - leu - gua - ade(pur) - pro ... his ... pyr ... trp. The reasons for peculiarities of the behaviour of transposons in Y. pestis bacteria are discussed.     

Cloning,expression, and crystallization of the V delta domain of a human gamma delta T-cell receptor.

T-lymphocytes recognize a wide variety of antigens through highly diverse cell-surface glycoproteins known as T-cell receptors (TCRs). These disulfide-linked heterodimers are composed of alpha and beta or gamma and delta polypeptide chains consisting of variable (V) and constant (C) domains non-covalently associated with at least four invariant chains to form the TCR-CD3 complex. It is well established that alpha beta TCRs recognize antigen in the form of peptides bound to molecules of the major histocompatibility complex (MHC); furthermore, information on the three-dimensional structure of alpha beta TCRs has recently become available through X-ray crystallography. In contrast, the antigen specificity of gamma delta TCRs is much less well understood and their three-dimensional structure is unknown. We have cloned the delta chain of a human TCR specific for the MHC class I HLA-A2 molecule and expressed the V domain as a secreted protein in the periplasmic space of Escherichia coli. Following affinity purification using a nickel chelate adsorbent, the recombinant V delta domain was crystallized in a form suitable for X-ray diffraction analysis. The crystals are orthorhombic, space group P2(1)2(1)2 with unit cell dimensions a = 69.9, b = 49.0, c = 61.6 A. and diffract to beyond 2.3 A resolution. The ability of a V delta domain produced in bacteria to form well-ordered crystals strongly suggests that the periplasmic space can provide a suitable environment for the correct in vivo folding of gamma delta TCRs.     

Photoelectric conversion by bacteriorhodopsin in charged synthetic membranes.

Photoelectroactivity of oriented purple membrane layers attached to an ion exchange film has been investigated. The action spectrum of the photocurrent followed the absorption spectrum of bacteriorhodopsin. The intactness of structure and function of bacteriorhodopsin was demonstrated by studies of absorption and photocycle kinetics. The direction of the photocurrent suggests that the extracellular surface of purple membrane is more positive. Photocurrents as high as 20 microA cm-2 were obtained in some preparations. The dependence of steady-state photocurrents on intensity of illumination and temperature was also studied. The initial rate of build-up of photocurrent depends linearly on the intensity of illumination while the off rate does not exhibit any dependence on the intensity of illumination. With rise in temperature an increase in the steady state photocurrent has been observed. This dependence was found to be linear when increase of the photocurrent due to proton translocation alone was considered.     

Possible mechanisms of the occurrence of chromosome aberrations. II. The formation of aberrations induced by UV irradiation

The report is concerned with one of possible mechanisms of emergence of chromosome aberrations after UV-irradiation of mammalian cells. The process is initiated by DNA cross-links following thymine dimerization, and is completed during mitosis. A model to account for formation of chromosome aberrations has been offered. It is compatible with the modern concept of a scaffolding model for metaphase chromosome structure in which the scaffold organizes DNA into loops along its length. The model predicts the importance of a process of mitotic chromosome isolation during aberration formation (in addition to the processes of DNA replication, reparation and chromosome association). Another feature of the model is an attempt to describe formation of aberrations under conditions of true DNA reparation.