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71.
Kinetic and allosteric propeties of highly purified "biosynthetic" L-threonine dehydratase from brewer's yeast S. carlbergensis were studied at three pH values, using L-threonine and L-serine as substrates. It was shown that the plot of the initial reaction rate (v) versus initial substrate concentrations ([S]0 pH 6.5 is hyperbolic (Km=5.0.10-2M), while these at pH 7.8 and 9.5 have a faintly pronounced sigmoidal shape with fast occurring saturation plateaus ([S]0.5= 1.0.10-2 and 0.9.10-2M, respectively). the ratios between L-threonine and L-serine dehydratation rates depend on pH. The kinetic properties and the dependence of substrate specificity on pH suggest that the enzyme molecule undergoes pH-induced (at pH 7.0) conformational changes. The determination of pK values of the enzyme functional groups involved in L-threonine binding demonstrated that these groups have pK is approximately equal to 7.5 and 9.5. The latter group was hypothetically identified as a epsilon-NH2-group of the lysine residue. High concentrations of the allosteric inhibitor (L-isoleucine) decrease the rates of L-threonine and L-serine dehydratation and induce the appearance (at pH 6.5) or increase (at pH 7.9 and 9.5) of homotropic cooperative interactions between the active sites in the course of L-threonine dehydratation. The enzyme inhibition by L-isoleucine increases with a decrease of L-threonine concentrations. Low L-isoleucine concentrations, as well as the enzyme activator (L-valine) stimulate the enzyme at non-saturating substrate concentrations (when L-threonine or L-serine are used as substrates) without normalization of (v) versus [S]0 plots. The maximal activation of the enzyme is observed at pHG 8.5--9.0. It is assumed that the molecule of "biosynthetic" L-threonine dehydratase from brewer's yeast contains two types of sites responsible for the effector binding, i.e., "activatory" and "inhibitory" ones. 相似文献
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Toyopearl media for gel filtration tend to adsorb proteins at high ionic strength, presumably by hydrogen bonding. This is used in the technique proposed here for resolution of crude protein mixtures and initial purification of their components. Proteins can be selectively adsorbed on a column of Toyopearl at high ammonium sulfate concentration and then eluted by decreasing the salt concentration. This single-step procedure can replace the usual salt fractionation of protein mixtures, which is demonstrated with yeast cytochrome oxidase. 相似文献
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Pomortsev A. A. Rubanovich A. V. Kovaleva O. N. Lyalina E. V. 《Russian Journal of Genetics》2019,55(11):1347-1359
Russian Journal of Genetics - Polymorphism of hordeins encoded by the Hrd A and Hrd B loci was studied using starch gel electrophoresis in 13 barley landrace accessions and in 31 wild barley... 相似文献
78.
Firstova A. A. Kofanov E. R. Zakshevskaya V. M. Kovaleva M. I. 《Russian Journal of Bioorganic Chemistry》2019,45(3):204-213
Russian Journal of Bioorganic Chemistry - A method for the high-yield synthesis of several amides of carboxylic acids containing an imide, a cyclohexene, and a norbornene cycle as well as fragments... 相似文献
79.
V. G. Artyukhov T. A. Kovaleva M. G. Kholyavka L. A. Bityutskaya M. V. Grechkina T. B. Obraztsova 《Biophysics》2009,54(6):675-680
It has been found using a combination of atomic force microscopy with infrared spectroscopy, gel chromatography, and electrophoresis
that inulinase from Kluyveromyces marxianus Y-303 has oligomeric structure, which includes two subunits differing in size, molecular mass, and catalytic activity. It
has been shown that the division of the inulinase dimer into monomers leads to an increase in the number of irregular regions
by 6% for subunit 1 (54.8 kDa) and by 10% for subunit 2 (8.4 kDa) compared with the native enzyme. 相似文献
80.
I. V. Kozhukharova I. I. Fridlyanskaya Z. V. Kovaleva N. A. Pugovkina L. L. Alekseenko V. V. Zenin K. M. Ivantsov O. K. Leont’eva T. M. Grinchuk N. N. Nikolsky 《Cell and Tissue Biology》2009,3(6):519-526
Novel human embryonic stem cell lines C612 and C910 have been established from atching blastocytes. Cells were cultivated in mTeSR medium on a mouse fibroblast feeder layer; they exhibit common pluripotent markers, such as alkaline phosphatase, Oct 3/4, SSEA-4, Nanog, Rex1. The immunophenotyping of these cells by flow cytometry revealed CD90 (Thy-1) and CD117 (c-kit) antigens and showed weak or no expression of CD13, CD34, CD45, CD130, and HLA class I and II antigens, which is typical for human embryonic stem cells. Karyotypic structure of C612 and C910 assayed by the G-banding of metaphase plates is normal in both chromosome number and structure. The cells generate embryoid bodies, undergo spontaneous differentiation, and express three germ-layer markers (nestin, keratin, vimentin ectoderm), α-fetoprotein (entoderm), muscle α-actinin (mesoderm), i.e., possess pluripotent potency. Thus, C612 and C910 display accepted human embryonic stem cell properties, including unlimited self-renewal, expression of pluripotent markers, ability to differentiate into three germ layers, and are diploid; therefore, they may be of potential use for fundamental research, as well as for replacement therapy studies. 相似文献