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1.
Synthesis in vitro of vesicular stomatitis virus proteins in cytoplasmic extracts of L cells 总被引:8,自引:0,他引:8
H P Ghosh F Toneguzzo S Wells 《Biochemical and biophysical research communications》1973,54(1):228-233
All five vesicular stomatitis virus (VSV) proteins, namely, L, G,N,NS and M are synthesized by a post-nuclear extract from cultured L cells infected with VSV. When, however, membrane bound polysomes are removed from the cytoplasmic extract only four virus specific proteins (L,N,NS and M) were synthesized. 相似文献
2.
Affinity chromatography has been used to isolate and compare the peanut agglutinin receptors from neuraminidase-treated human, bovine and porcine erythrocyte membranes. Passage of Triton X-100-solubilised membrane material through either Sepharose- or acrylamide-based affinity columns resulted in the reversible binding of receptor molecules to the immobilised lectin. Elution with 0.2M galactose released specifically bound glycoprotein fractions, the composition and molecular weights of which were determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate.Carbohydrate analysis by gas chromatography identified these bound glycoprotein fractions as the major sources of the disaccharide in these membranes. It is suggested that these isolated fractions represent a discrete population of glycoproteins within the membranes studied, which possess both - and carbohydrates. 相似文献
3.
Pamela Stanley Grace Vivona Paul H. Atkinson 《Archives of biochemistry and biophysics》1984,230(1):363-374
Carbohydrate moieties derived from the G glycoprotein of Vesicular Stomatitis Virus (VSV) grown in parental Chinese hamster ovary (CHO) cells and the glycosylation mutant Lec4 have been analyzed by high-field 1H NMR spectroscopy. The major glycopeptides of and were purified by their ability to bind to concanavalin A-Sepharose. The carbohydrates in this fraction are of the biantennary, complex type with heterogeneity in the presence of α(2,3)-linked sialic acid and α(1,6)-linked fucose residues. A minor glycopeptide fraction, which does not bind to concanavalin A-Sepharose but which binds to pea lectin-agarose, was also investigated by 1H NMR spectroscopy. These carbohydrates are complex moieties which appear to contain N-acetylglucosamine in β(1,6) linkage. Their spectral properties are most similar to those of a triantennary complex oligosaccharide containing a 2,6-disubstituted mannose α(1,6) residue. Carbohydrates of this type are not found among the glycopeptides of VSV grown in the Lec4 CHO glycosylation mutant. 相似文献
4.
Thermal denaturation of nucleocapsids of wild type (WT) vesicular stomatitis virus (VSV), containing only the nucleocapsid protein (N) and viral RNA, caused a “melting” that resulted in an A260nm absorbance increase of 140%. The nucleocapsids of two temperature-sensitive () VSV mutants, G31BP and G22, both underwent larger absorbance increases of 251% and 177% respectively, suggesting these nucleocapsids are complexed by weaker N protein: RNA interactions than the WT-VSV. Two other mutants, G31 and G41 underwent A260nm increases either similar to, or smaller than, that measured with WT-VSV nucleocapsids. RNA synthesis by G31BP in infected cells was also found to be decreased at elevated temperatures. This temperature sensitive defect in viral RNA metabolism in G31BP may be the result of weaker protein:RNA interactions associated with the nucleocapsid. 相似文献
5.
M. Thirunavukkarasu Fergus G. Priest 《Biochemical and biophysical research communications》1983,114(2):677-683
α-Glucosidase was membrane bound during exponential growth of but was released into the medium during stationary phase. It could be partially removed from exponential phase cells by washing with NaCl (0.5 M). α-Amylase was exclusively extracellular and could not be detected in cells. Polysomes were prepared from exponential phase cells and separated into membrane bound and soluble fractions. chain completion and immunoprecipitation showed that α-glucosidase and α-amylase were synthesized by membrane bound and not by soluble ribosomes. 相似文献
6.
Expression and subcellular distribution of mouse cytochrome P1-450 mRNA as determined by molecular hybridization with cloned P1-450 DNA 总被引:1,自引:0,他引:1
Cytochrome P1-450 (P1-450) is defined as that cytochrome most closely associated with 3-methylcholanthrene (MC)-induced aryl hydrocarbon hydroxylase (AHH) activity. Recently a cloned DNA sequence (clone 46) was shown to represent a portion of the P1-450 structural gene [Negishi : 800–804 (1981)]. Poly(A+)-enriched RNA was isolated from total liver homogenate, membrane-bound polysomes and from free polysomes at various times after MC treatment of -responsive (B6) and -nonresponsive (D2) inbred mice. The poly(A+)-enriched RNA was separated by methylmercury-agarose gel electrophoresis and hybridized to nick-translated [32P]DNA from clone 46. By means of this RNA-DNA hybridization, only 6% of total polysomal P1-450 mRNA exists in free polysomes after 24 h of MC treatment. The data indicate that the endoplasmic reticulum is the principal site of synthesis for this integral microsomal protein.Studies of induction kinetics following MC treatment provided the evidence of the rapid increase of total liver and membrane bound P1-450 mRNA preceding the synthesis of apo-P1-450 and the increase of AHH activity. 相似文献
7.
Phenol extraction of bovine milk fat globule membrane gave a glycoprotein fraction which, in sodium dodecyl sulphate electrophoresis, showed three major bands, all staining for both protein and carbohydrate. Alkaline borohydride treatment and desialylation of the glycoprotein fraction released the reduced disaccharide β-d-galactosyl(1 → 3)- (T-antigen), which was identified by gas chromatography using a standard. All of the disaccharide units in the native glycoprotein were shown to be substituted by sialic acid, and a tetrasaccharide containing the disaccharide plus two molecules of sialic acid was isolated following alkaline borohydride treatment of the glycoprotein and gel filtration. Periodate oxidation of native and desialylated glycoprotein, together with paper chromatography of alkali degraded oligosaccharide fragments, indicated that the major alkali-labile oligosaccharide of the glycoprotein fraction is a tetrasaccharide containing substituted by sialic acid at position C3 of the galactosyl and position C6 of the residue. Evidence was also obtained for the presence of small amounts of unsubstituted alkali-labile linked directly to protein in the native glycoprotein.Serological evidence using agglutinins from Vicia graminea, Arachis hypogoea and human anti-T serum confirmed the presence in the native glycoprotein of a sialic acid substituted T-antigen. Similar evidence using agglutinins from Helix pomatia and Cepaea hortensis also confirmed the presence of terminal alkali-labile in the native glycoprotein. 相似文献
8.
Sergio L. Abreu F.Carter Bancroft 《Biochemical and biophysical research communications》1978,82(4):1300-1305
Human diploid fibroblast (FS-4) cells were induced to produce interferon mRNA by exposure to poly(rI)·poly(rC) plus cycloheximide. The intracellular location of interferon mRNA was investigated by differential centrifugation of the cytoplasm into a membrane (pellet) and a free (supernatant) fraction, followed by injection of mRNA isolated from either fraction into oocytes. When translation in FS-4 cells was prevented, most (85–90%) of the interferon mRNA activity was found in the free fraction. However, when translation was permitted, most (80–95%) of the interferon mRNA activity was found in the membrane fraction. These results are consistent with the predictions of the “signal hypothesis” (Blobel and Dobberstein, J. Cell Biol. 1975, :835) for secretory proteins. 相似文献
9.
Lionel Lelievre Alain Zachowski Daniele Charlemagne Patrice Laget Alain Paraf 《生物化学与生物物理学报:生物膜》1979,557(2):399-408
Treatment of plasma membrane isolated from murine plasmocytoma MOPC 173 with an EDTA-containing buffer resulted in a 300-fold increase in sensitivity of Mg2+-ATPase to ouabain. This phenomenon was associated with the solubilization by EDTA of phospholipid free proteins (approx. 30 000–34 000 daltons) from the cytoplasmic face of the plasma membrane and with removal of about 90% of the membrane bound Ca2+. The recovery of the original resistance to ouabain required specifically Ca2+ and was associated with a binding of the solubilized proteins to the membrane. 相似文献
10.
Cytosine-rich messenger RNA from carrot root discs 总被引:2,自引:0,他引:2
D A Stuart T J Mozer J E Varner 《Biochemical and biophysical research communications》1982,105(2):582-588
The total poly A+ RNA from aerated carrot root discs was further fractionated into a cytosine-rich mRNA fraction by oligo (dG) cellulose chromatography. C-rich mRNA was purified at least 10-fold by this procedure and, when translated in wheat germ lysates, codes for 57 and 53 Kdalton peptides. Translation in double label amino acid mixtures indicates that C-rich mRNA codes for proline-rich peptides which may be precursors to the hydroxyproline-rich glycoprotein synthesized by this tissue. 相似文献