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91.
beta-Br-ethylamine is both a substrate and an irreversible inhibitor of amine oxidase from Aspergillus niger. The enzyme catalyzes the nonoxidative elimination of HBr from beta-Br-ethylamine to form acetaldehyde. beta-Br-ethylamine meets several criteria for an irreversible substrate analog or suicide inhibitor. 1) It inactivates the oxidized enzyme, but not the reduced enzyme. 2) The Michaelis constant for beta-Br-ethylamine in the elimination reaction showed a similar magnitude to that of the related constant found when the haloamine acted as an inhibitor. 3) The enzyme was protected from the inactivation by the co-existence of the substrate. 4) Inactivation with beta-Br-[14C]ethylamine resulted in the incorporation of radioactivity corresponding to 1 mol of the label/mol of the monomeric unit of the enzyme and a decrease of 1 mol of the -SH group. 5) Inactivation was accompanied by the formation of a new absorption peak at 320 nm which was bleached by addition of NaBH4. 相似文献
92.
Antibodies introduced into living cells by red cell ghosts are functionally stable in the cytoplasm of the cells. 总被引:10,自引:0,他引:10
The function and fate of antibodies introduced into living cells by red cell ghosts were studied using CRM 176 (a mutant diphtheria toxin having lower toxicity than the wild-type) and antibody against fragment A of diphtheria toxin. IgG labeled with iodine and FITC was found in the cytoplasm of the recipient cells. When about 1500 molecules of anti-fragment A antibody (rabbit IgG) were introduced into diphtheria toxin-sensitive Vero cells or FL cells, these cells became resistant to the toxin and formed normal colonies. It was calculated from the survival of cells without anti-fragment A IgG under these conditions that about 300 molecules of fragment A-176 were transferred to the cells. These results showed that the antigen-antibody reaction took place in living cells as effectively as in a cell-free system. The functional stability of antibody IgG in cells was examined by exposing Vero cells containing a subminimal amount of anti-fragment A IgG (about 1000 molecules) to the toxin for 2 hr at various times after the introduction of anti-fragment A IgG. More than 50% of the initial activity of the antibody to neutralize toxin still remained even after incubation of the cells at 37°C for 20 hr. The same degree of stability was also demonstrated using iodine-labeled specific anti-fragment A IgG. The IgG recovered from the recipient cells after various times of incubation at 37°C retained its full ability to bind to fragment A-conjugated Sepharose 4B, although the total amount of IgG associated with the cells decreased about 50% in 24 hr. 相似文献
93.
K. Murata T. Uchida K. Tani J. Kato I. Chibata 《Applied microbiology and biotechnology》1979,7(1):45-51
Summary Whole cells of Achromobacter butyri OUT 8004 having polyphosphate glucokinase activity were immobilized in polyacrylamide gel. The immobilized cells were activated by organic solvents, especially acetone. The immobilization resulted in increased stability of polyphosphate glucokinase. Continuous high yield production of G-6-P from glucose and metaphosphate was performed with an immobilized cell column, which had a half-life of approximately 20 days.Abbreviations G-6-P
glucose-6-phosphate
- G-1-P
glucose-1-phosphate
- Cation-S
stearyl trimethyl ammonium chloride
- SDS
sodium dodecyl sulfate
- Tris
tris(hydroxymethyl)-aminomethane; p-NPP, p-nitrophenyl phosphate
- S.V.
space velocity 相似文献
94.
Summary The existence of the surface-connected canalicular system (SCCS) has been demonstrated in semithick sections of the frog thrombocytes by the use of a high voltage electron microscope. The SCCS of the thrombocytes in Rana catesbeiana and Rana nigromaculata consists of numerous canaliculi and vesicles with a diameter of 250 nm, which join with one another to make a complex network throughout the cytoplasm. Although the SCCS of Xenopus laevis fits well into the pattern described in Rana catesbeiana, the diameter of the canaliculi of the SCCS is about 500 nm. The results of this study suggest that the SCCS is a specific organelle of the thrombocyte system common to submammals and mammals. 相似文献
95.
Appearance of spore coat protein in the cell extracts of Bacillus subtilis asporogenic mutants. 下载免费PDF全文
By use of the antigen-antibody techniques we have studied whether asporogenic mutants of Bacillus subtilis can synthesize the spore coat protein. Antibody specific to spore coat protein was prepared and used to demonstrate that the spore coat protein was synthesized at the early stage of sporulation. We report here that asporogenic mutants synthesize the spore coat protein. 相似文献
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99.
The Gc types in the Japanese 总被引:3,自引:0,他引:3
100.