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891.
Free energy perturbation calculations on binding and catalysis after mutating Asn 155 in subtilisin 总被引:1,自引:0,他引:1
Site-directed mutagenesis is a very powerful approach to altering the biological functions of proteins, the structural stability of proteins and the interactions of proteins with other molecules. Several experimental studies in recent years have been directed at estimating the changes in catalytic properties, (rates of binding and catalysis) in site-directed mutants of enzymes compared to the native enzymes. Simulation approaches to the study of complex molecules have also become more powerful, in no small measure owing to the increase in computer power. These simulations have often allowed results of experiments to be rationalized and understood mechanistically. A new approach called the free-energy pertubation method, which uses statistical mechanics and molecular dynamics can often be used for quantitative calculation of free energy differences. We have applied such a technique to calculate the differential free energy of binding and free energy of activation for catalysis of a tripeptide substrate by native subtilisin and a subtilisin mutant (Asn 155----Ala 155). Our studies lead to a calculated difference in free energy of binding which is relatively small, but a calculated change in free energy of catalysis which is substantial. These energies are very close to those determined experimentally (J. A. Wells and D. A. Estell, personal communication), which were not known to us until the simulations were completed. This demonstrates the predictive power and utility of theoretical simulation methods in studies of the effects of site-specific mutagenesis on both enzyme binding and catalysis. 相似文献
892.
The binding of phosphorylated heavy meromyosin to regulated F-actin in ghost fibers at high Ca2+ concentration increases, and at low Ca2+ concentration decreases, the anisotropy of intrinsic tryptophan fluorescence of F-actin. The effect is opposite to the effect of the binding of dephosphorylated heavy meromyosin. 相似文献
893.
894.
Synthesis and some characteristics of [13C]-specially enriched tetragastrin and the related compound
[13C]-enriched tetragastrin and the related compound were synthesized in solution. Conversion of S-[13C]methylated tetragastrin to the enriched tetragastrin gave 10.5 ppm upfield chemical shift of Cepsilon resonance. The potency of the synthetic tetragastrin to stimulate gastric acid secretion was virtually identical with that of pentagastrin (ICI). 相似文献
895.
The N-terminal fragments (residues 1-51 and 1-59) obtained by selective tryptic cleavage of native lac repressor retain the ability to bind DNA. These fragments (headpieces) are monomeric and form complexes which resemble those of tetrameric repressor with non-operator DNA. But, they do not show the high specificity of repressor for operator sequences. The DNA binding has been demonstrated by filter-binding assay as well as in solution using absorption, circular dichroism, and fluorescence measurements. 相似文献
896.
897.
Epstein-Barr virus genome-positive T lymphocytes in a boy with chronic active EBV infection associated with Kawasaki-like disease 总被引:21,自引:0,他引:21
H Kikuta Y Taguchi K Tomizawa K Kojima N Kawamura A Ishizaka Y Sakiyama S Matsumoto S Imai T Kinoshita 《Nature》1988,333(6172):455-457
Epstein-Barr virus (EBV), a ubiquitous human herpesvirus and an aetiological agent of infectious mononucleosis, has a unique tropism for B lymphocytes. Clinical and laboratory features of chronic active EBV infections are chronic or persistent infectious mononucleosis-like symptoms and high antibody titre against early antigens (EA). Kawasaki disease (KD), aetiology unknown, is thought to be self-limited immunologically mediated vasculitis. Clinical features of KD are fever, rash, mucositis, lymphadenopathy and coronary artery damage. We report here a child with chronic active EBV infection accompanied by dilatation of coronary arteries. All the EBV-determined nuclear antigen (EBNA)-positive lymphocytes had exclusively CD4 antigen, as revealed by dual staining immunofluorescence analysis. Southern blot hybridization showed that the purified CD4+ cells harboured EBV genome. 相似文献
898.
D. Galdean D. Petrasincu P. Alangiu S. Ibric N. Voiculetz 《Cellular and molecular life sciences : CMLS》1986,42(5):572-574
Summary The protective effect of Thiola against the genotoxicity, induced by benzo(a)pyrene, in vitro and in vivo, was investigated. By association of Thiola to benzo(a)pyrene a significant decrease of the numerical and structural chromosome aberrations and a reduction of the incidence of c-mitoses has been obtained in human diploid cells, i.e. human embryonic lung fibroblasts of the cell-line ICP-23, and C56Bl/6 mouse bone marrow cells. 相似文献
899.
M. E. Hoffmann D. B. Ciampi N. Duran 《Cellular and molecular life sciences : CMLS》1987,43(2):217-220
Summary A radical anion of 2,3,5,6-tetrahydroxy-2,5-cyclohexadiene-1,4-dione was detected using the EPR technique by complexing with Zn ions. Hydrogen peroxide, superoxide radical anion and hydroxyl radical were also detected in the reaction mixture. Kinetic study and product distribution indicated a probable mixed type one- and two-electron transfer mechanism. A possible relationship between the autooxidation process and the biological activity of substituted quinones was suggested. 相似文献
900.
Regulation of transcription. Flexible interpretation 总被引:1,自引:0,他引:1