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Membrane-associated phosphoinositidase C activity has been identified in Dictyostelium discoideum using phosphatidylinositol 4,5-bisphosphate as exogenous substrate. Maximal activity was observed with 0.4 mM phosphatidylinositol 4,5-bisphosphate at pH 7.0. The enzyme was stimulated
by micromolar concentrations of free calcium with maximal activity at 100 μM. 相似文献
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Mapping the active site of yeast RNA polymerase B (II) 总被引:11,自引:0,他引:11
M Riva C Carles A Sentenac M A Grachev A A Mustaev E F Zaychikov 《The Journal of biological chemistry》1990,265(27):16498-16503
Yeast RNA polymerase B (II) was incubated with a collection of 13 different nucleotide derivatives and affinity labeled by allowing DNA-directed phosphodiester bond formation. The 32P-labeled site was localized in the C-terminal part of the B150 subunit by microsequencing a proteolytic fragment, then further mapped by a combination of extensive or single-hit chemical cleavage reactions and analysis of the labeled peptide patterns. The affinity label was mapped to between Asn946 and Met999, within one of the nine regions that are conserved between B150 and the bacterial beta subunit. The results underscore the conservative evolution of the catalytic center of eukaryotic and bacterial RNA polymerases. 相似文献
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Active site labeling of the RNA polymerases A, B, and C from yeast 总被引:13,自引:0,他引:13
M Riva A R Sch?ffner A Sentenac G R Hartmann A A Mustaev E F Zaychikov M A Grachev 《The Journal of biological chemistry》1987,262(30):14377-14380
RNA polymerases A, B, and C from yeast were modified by reaction with 4-formylphenyl-gamma-ester of ATP as priming nucleotide followed by reduction with NaBH4. Upon phosphodiester bond formation with [alpha-32P]UTP, only the second largest subunit, A135, B150, or C128, was labeled in a template-dependent reaction. This indicates that these polypeptide chains are functionally homologous. The product covalently bound to B150 subunit was found to consist of a mixture of ApU and a trinucleotide. Enzyme labeling exhibited the characteristic alpha-amanitin sensitivity reported for A and B RNA polymerases. Labeling of both large subunits of enzyme A and B but not of any of the smaller subunits was observed when the reduction step stabilizing the binding of the priming nucleotide was carried out after limited chain elongation. These results illustrate the conservative evolution of the active site of eukaryotic RNA polymerases. 相似文献
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Shashkov Alexander S. Tul’skaya Elena M. Potekhina Natalia V. Dmitrenok Andrey S. Senchenkova Sofia N. Zaychikov Vlad A. Dorofeeva Lubov V. Evtushenko Lyudmila I. 《Biochemistry. Biokhimii?a》2021,86(4):506-516
Biochemistry (Moscow) - Rathayibacter sp. VKM Ac-2759 (family Microbacteriaceae, class Actinobacteria) contains two glycopolymers in the cell wall. The main chain of rhamnan, glycopolymer 1, is... 相似文献
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T G Maksimova A A Mustayev E F Zaychikov D L Lyakhov V L Tunitskaya A Kh Akbarov S V Luchin V O Rechinsky B K Chernov S N Kochetkov 《European journal of biochemistry》1991,195(3):841-847
A highly selective affinity labeling of T7 RNA polymerase with the o-formylphenyl ester of GMP and [alpha-32P]UTP was carried out. The site of the labeling was located using limited cleavages with hydroxylamine, bromine, N-chlorosuccinimide and cyanogene bromide and was identified as the Lys631 residue. Site-directed mutagenesis using synthetic oligonucleotides was used to substitute Lys631 by a Gly, Leu or Arg residue. Kinetic studies of the purified mutant enzymes showed alterations of their polymerizing activity. For the Lys----Gly mutant enzyme, anomalous template binding was observed. 相似文献
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Zaychikov V. A. Potekhina N. V. Dmitrenok A. S. Fan Ding Tul’skaya E. M. Dorofeeva L. V. Evtushenko L. I. 《Microbiology》2021,90(3):343-348
Microbiology - Presence of the cell wall glycopolymer rhamnan was established for members of the genus Curtobacterium (family Microbacteriaceae) by using chemical and NMR spectroscopic methods. The... 相似文献
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