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51.
Groups of CBA mice were administered [35S] methionine (1 mCi/mouse). Non-histone proteins, H1 and H10 histones and nucleosomal core histones were isolated from different issues by selective extractions. The measurements of radioactivity of individual bands and autoradiography of dry gels were used to identify methionine-containing and methionine-free histone variants. H1A and H1B histone variants extracted with 5% perchloric acid were methionine-free. However, minor sub-fractions of these histones which are more tightly bound to DNA (and which can be extracted only with 0.25 N HC1) contained [35S] methionine and did show a higher specific activity than methionine-containing nucleosomal hitones. Cyanogen Bromide reaction which destroys non-histone proteins and methionine-containing nucleosomal histones removes radioactivity but does not alter the position of methionine-containing H1 minor bands. This indicates that the radioactive methionine occupies only the N-terminus of the H1 molecules. It is suggested that this methionine is an uncleaved initiator methionine. The presence of these methionine-containing minor H1 subfractions varies in different tissues. 相似文献
52.
R. Ayola D. F. Condorelli N. Ragusa M. Renis M. Alberghina A. M. Giuffrida Stella Abel Lajtha 《Neurochemical research》1988,13(4):337-342
In vivo protein synthesis rates in various brain regions (cerebral cortex, cerebellum, hippocampus, hypothalamus, and striatum) of 4-, 12-, and 24-month-old rats were examined after injection of a flooding dose of labeled valine. The incorporation of labeled valine into proteins of mitochondrial, microsomal, and cytosolic fractions from cerebral cortex and cerebellum was also measured. At all ages examined, the incorporation rate was 0.5% per hour in cerebral cortex, cerebellum, hippocampus, and hypothalamus and 0.4% per hour in striatum. Of the subcellular fractions examined, the microsomal proteins were synthesized at the highest rate, followed by cytosolic and mitochondrial proteins. The results obtained indicate that the average synthesis rate of proteins in the various brain regions and subcellular fractions examined is fairly constant and is not significantly altered in the 4 to 24-month period of life of rats.A preliminary report of these results was previously presented at: WFN-ESN Joint Meeting on: Cerebral Metabolism in Aging and Neurological Disorders, Baden, August 28–31, 1986. 相似文献
53.
54.
The addition of antimycin A during the logarithmic phase of growth of heterotrophic Euglena gracilis cultures (in lactate or glucose medium) was immediately followed by decreased respiration and a cessation of grwoth. Induced cyanideresistent respiration appeared 5 h after the addition of the inhibitor then the cells started to grow again and could be cultured in the presence of antimycin A. Thus the cells exhibited a cyanide-and antimycin-resistant respiration which was, in addition, sensitive to salicylhydroxamic acid and propylgallate. Antimycin-adapted Euglena and control cells were compared for their biomass production and protein synthesis. The difference in growth yield between control and antimycin-adapted cells was not as high as would be expected if only the first phosphorylation site of the normal respiratory chain was active in the presence of antimycin A. Furthermore, the ability to incorporate labelled valine into proteins, under resting-cell conditions, was not changed. Strong correlations were established between the effects of respiratory effectors on O2 consumption and valine incorporation. These results suggest that sufficient energy for protein synthesis and growth is provided by the operation of the cyanide-resistant respiratory pathway in antimycin-adapted Euglena.Abbreviations DNP
dinitrophenol
- PG
propylgallate
- SHAM
salicylhydroxamic acid 相似文献
55.
56.
Glyoxysomal citrate synthase (gCS) was purified from crude extracts of watermelon (Citrullus vulgaris Schrad.) cotyledons, yielding a homogenous protein with a subunit MW of 48 kDa. The enzyme was selectively inhibited by 5,5-dithiobis-(2-nitrobenzoic acid), allowing quantification in the presence of the mitochondrial isoenzyme (mCS). Differences were also observed with respect to inhibition by ATP (k
i=2.6 mmol · l-1 for gCS, k
i=0.33 mmol · l-1 for mCS). The antibodies prepared against gCS did not cross-react with mCS. The immunocytochemical localization of gCS by the indirect protein A-gold procedure was restricted to the glyoxysomal membrane or the peripheral matrix of glyoxysomes. Other compartments, e.g. the endoplasmic reticulum, were not labeled. Xenopus oocytes were used for the translation of watermelon polyadenylated RNA (poly(A)+RNA). A translation product with a MW of 51 kDa was immunoprecipitated by the anti-gCS antibodies. It was absent in controls without poly(A)+RNA or with preimmune serum. A similar translation product was also immunoprecipitated after cell-free synthesis of watermelon poly(A)+RNA in a reticulocyte system, in contrast to the in-vivo labeled gCS (48 kDa). It was concluded that gCS is synthesized as a higher-molecular-weight precursor.Abbreviations DTNB
5,5-dithiobis-(2-nitrobenzoic acid)
- gCS
glyoxysomal citrate synthase
- gMDH
glyoxysomal malate dehydrogenase
-
k
i
inhibitor constant
- mCS
mitochondrial citrate synthase
- OAA
oxaloacetate
- poly(A)+RNA
polyadenylated RNA
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis 相似文献
57.
Immunogold labelling was used to study the distribution of acyl carrier protein (ACP) in Escherichia coli and a variety of plant tissues. In E. coli, ACP is distributed throughout the cytoplasm, confirming the observation of S. Jackowski et al. (1985, J. Bacteriol., 162, 5–8_. In the mesocarp of Avocado (Persea americana) and maturing seeds of oil-seed rape (Brassica napus cv. Jet Neuf), over 95% of the ACP is localised to plastids. The protein is almost exclusively located in the chloroplasts of leaf material from oil-seed rape. Approximately 80% of the gold particles associated with the ACP were further localized to the thylakoid membrane of the chloroplast. Since acetyl-CoA carboxylase has been reported to be localized to the thylakoid membrane (C.G. Kannangara and C.J. Jensen, 1975, Eur. J. Biochem., 54, 25–30), these results are consistent with the view that the two sequential enzymes in fatty-acid synthesis are in close spacial proximity.Abbreviations ACC
acetyl CoA carboxylase
- ACP
acyl carrier protein
- FAS
fatty-acid synthetase 相似文献
58.
王钧 《分子细胞生物学报》1988,(2)
pTiAch 5 TR区与在植物中积累agropine相关的三个基因片段已导入pINIIA或pACYC184,在大肠杆菌微细胞中表达出杂合蛋白。三个基因片段的pINIIA重组质粒的表达得到稳定蛋白、不稳定蛋白和在两个相表达出相似蛋白的三种结果。基因2'HindⅢ片段上的UGAA序列是在两个相表达相似蛋白的原因。从产生的杂合蛋白分子量和读码方式看,pTiAch 5中基因2'和基因1'的结构与pTi 15955近似,但由基因0'片段产生的杂合蛋白分子量比根据pTi15955 DNA序列数据推测的稍大。从表达强度和微细胞操作看,这个系统还不适应从大肠杆菌中方便地纯化杂合蛋白的需要。 相似文献
59.
Spyridon Vamvakas Wolfgang Dekant Dietmar Schiffmann Dietrich Henschler 《Cell biology and toxicology》1988,4(4):393-403
S-(chloroethyl)-cysteine (CEC) and S-(1,2-dichlorovinyl)cysteine (DCVO) have been proposed as intermediates in the metabolic transformation of the carcinogens 1,2-dichloroethane and 1,1,2-trichloroethylene. We have tested the ability of CEC and DCVC to induce DNA repair and genotoxic effects at the chromosomal level by comparative assessment of unscheduled DNA synthesis induction and micronucleus formation in Syrian hamster embryo fibroblasts. CEC induced a potent and dose-dependent response in both assays, whereas DCVC treatment resulted in a comparatively weak induction of DNA repair and failed to raise micronucleus formation above control rates. Inhibition of cysteine conjugate \gB-lyase diminished the effect of DCVC, but had no influence on the genotoxicity of CEC either in the unscheduled DNA synthesis or micronucleus assay.Abbreviations AOAA
aminooxyacetic acid
- CEC
S-(chloroethyl)-cysteine; \gB-lyase, cysteine conjugate -lyase
- DCE
1,2-dichloroethane
- DCVC
S(1,2-dichlorovinyl)-cysteine
- GSH
glutathione
- HU
hydroxyurea
- IBR
IBR-modified Dulbecco's Eagle's reinforced medium
- MN2
micronuclei/2,000 cells
- 4-NQO
4-nitroquinoline-1-oxide
- SHE
Syrian hamster embryo fibroblasts; 3H-Thd, 3H-thymidine
- TCE
1,1,2-trichloroethylene
- UDS
unscheduled DNA synthesis 相似文献
60.
Richard C. Woodman John T. Curnutte Bernard M. Babior 《Free radical biology & medicine》1988,5(5-6):355-361
We examined the effects of the recombinant human colony stimulating factors GM-CSF and G-CSF, cycloheximide (a protein synthesis inhibitor) and dihydrocytochalasin B (a microfilament disrupting agent) upon FMLP (N-formyl-methionyl-leucylphenylalanine)-stimulated O2 − production by neutrophils. We confirmed a time dependent augmentation of O2 − production following preincubation of neutrophils either alone or with colony stimulating factors. Furthermore, we found that GM-CSF, but not G-CSF, increased O2 − production at some concentrations of the stimulus. Preincubation of neutrophils with cycloheximide in the absence of CSF caused a marked fall in O2−-production that was first evident at 2 hours. The fall in O2−-forming capacity caused by cycloheximide was much less pronounced if dihydrocytochalasin B was also included in the preincubation buffer. These findings suggest a previously unrecognized role for de novo protein synthesis in maintaining the ability of neutrophils to manufacture O2−, and support earlier studies indicating that the cycling of FMLP receptors between the cell membrane and an intracellular compartment is important in determining the magnitude of the respiratory burst in FMLP-stimulated neutrophils. 相似文献