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11.
M Laloux E Van Schaftingen J Francois H G Hers 《European journal of biochemistry》1985,148(1):155-159
Experiments performed at micromolar concentrations of inorganic phosphate support the conclusion that liver phosphofructokinase 2 would be completely inactive in the absence of inorganic phosphate or arsenate. The concentration of inorganic phosphate that allowed half-maximal activity decreased with increasing pH, being approximately 0.11 mM at pH 6.5 and 0.05 mM at pH 8. The effect of phosphate was to increase V and to decrease Km for fructose 6-phosphate, without affecting Km for ATP. Citrate and P-enolpyruvate inhibited the enzyme non-competitively with fructose 6-phosphate and independently of the concentration of inorganic phosphate. Phosphorylation of the enzyme by the catalytic subunit of cyclic-AMP-dependent protein kinase did not markedly modify the phosphate requirement and its effect of inactivating phosphofructokinase 2 could not be counteracted by excess phosphate. A nearly complete phosphate dependency was also observed with phosphofructokinase 2 purified from Saccharomyces cerevisiae or from spinach leaves. By contrast, the fructose 2,6-bisphosphatase activity of the liver bifunctional enzyme was not dependent on the presence of inorganic phosphate. Phosphate increased this activity about threefold when measured in the absence of added fructose 6-phosphate and a half-maximal effect was reached at approximately 0.5 mM phosphate. Like glycerol phosphate, phosphate counteracted the inhibition of fructose 2,6-bisphosphatase by fructose 6-phosphate, but a much higher concentration of phosphate than of glycerol phosphate was required to reach this effect. 相似文献
12.
J Francois 《Biochimie》1985,67(9):1035-1042
The collagen from the mesenteric sheath of the tenebrionid insect Tenebrio molitor was extracted by limited pepsin digestion and purified. This collagen was characterized using CM-cellulose chromatography, sodium-dodecylsulfate disc-gel electrophoresis and aminoacid analysis. This molecule was found to be assembled from three identical alpha chains and could be represented by the formula (alpha) 3. The amino acid composition is characteristic of collagen (one-third glycine, high iminoacid content), with high content of hydroxylysine and low content of alanine. Cyanogen bromide digests of these chains indicated that they are not related to any of the known invertebrate or vertebrate chains of interstitial collagens. The molecular weight (M = 280000D) and length (290 nm) were typical, and the banding patterns of the segment-long-spacing crystallites (SLS) and of the reconstitued fibrils were very similar to type I collagen. The denaturation temperature (Td) was 30.7 degrees C and correlated with the total pyrrolidine content as observed in other collagens (von Hippel & Wong's relation). It was concluded that the collagen from this insect showed the classical biochemical and biophysical features of other invertebrate interstitial "primitive" collagens. 相似文献
13.
Jef D. Boeke Francois La Croute Gerald R. Fink 《Molecular & general genetics : MGG》1984,197(2):345-346
Summary Mutations at the URA3 locus of Saccharomyces cerevisiae can be obtained by a positive selection. Wild-type strains of yeast (or ura3 mutant strains containing a plasmid-borne URA3
+ gene) are unable to grow on medium containing the pyrimidine analog 5-fluoro-orotic acid, whereas ura3
– mutants grow normally. This selection, based on the loss of orotidine-5-phosphate decarboxylase activity seems applicable to a variety of eucaryotic and procaryotic cells. 相似文献
14.
Jojoba plants of the selection ‘Vista’ were grown for two years in the greenhouse at four levels of soil salinity. All plants reached anthesis with no symptom of major injury because of the treatments. The following changes were noted on treated plants: Leaves were thicker, with larger secretory and palisade cells. They had fewer stomata per unit area, smaller vascular cylinder and vascular cells, and discontinuous layers of secretory cells. Stems were thinner, with smaller pith diameter, smaller pith cells, and smaller vessel elements. Both leaves and stems had a higher moisture content. A higher accumulation of Cl and Na ions was observed in both leaves and stems. Mg ions decreased in leaves but increased in stems. Ca ions increased in leaves only. 相似文献
15.
16.
1. The separation of the alpha-components of codfish-skin and calf-skin collagen from the higher-molecular-weight components was accomplished by gel filtration with Bio-Gel P-300 resin in 5m-lithium chloride and at pH7.4. 2. From this fraction three distinct alpha-components, corresponding to the alpha1-, alpha2- and alpha3- chains of collagen, were isolated by free-flow electrophoresis in 6m-urea at 4 degrees . At the temperatures and pH values used there was no detectable degradation of the alpha-chains. 相似文献
17.
The present study depicts the presence of a gradient in the morphological characters ofThymelaea hirsuta (L.)Endl. leaves which correlated with the environmental gradient prevailing in the Western Mediterranean region of Egypt. The less arid and more calcareous habitats harbour individuals with obtuse and gentle curved leaf apices and gentle involute leaf margins. With the increase of aridity and decrease of CaCO3, the leaf apices become acute and strongly curved, and the leaf margins become strongly involuted. Significant variations in seed weight, seedling emergence and viability of seed embryos inT. hirsuta, in relation to habitat types, are also shown and discussed. 相似文献
18.
Joelle Vilamitjana-Amedee Reine Bareille Francois Rouais Arnold I. Caplan Marie-Francoise Harmand 《In vitro cellular & developmental biology. Animal》1993,29(9):699-707
Summary This study reports the selection and characterization of osteogenic precursors from human bone marrow which were isolated
by two “clonings” and successive subculturing. These cell lines express alkaline phosphatase activity. Gel electrophoresis
of [3H]-proline labeled cultures showed that the cloned cells produce only type I collagen. They synthetize osteocalcin and osteonectin.
They respond to 1,25 dihydroxy vitamin D3 by increasing osteocalcin synthesis and secretion, and to parathyroid hormone by increasing cyclic AMP synthesis. After the
third subculture in the absence of β-glycerophosphate, these cell lines formed lots of clusters which exhibit high alkaline
phosphatase activity and positive von Kossa staining. X-ray energy spectrum shows that these cells are surrounded by “budding”
structures containing calcium and phosphorus with a ratio Ca:P identical to those of pure hydroxyapatite. This process was
associated with45Ca uptake into the cells. All these data support the selection of osteogenic cells which may be of considerable clinical importance. 相似文献
19.
Sujoy Ghosh John W. Kyle Sara Dastgheib Francois Daussin Zhixiong Li Subhash Basu 《Glycoconjugate journal》1995,12(6):838-847
A 1-3 galactosyltransferase (GalT-3; UDP-Gal; GM2 1-3galactosyltransferase) was purified over 5100-fold from 19-day-old embryonic chicken brain homogenate employing detergent solubilization, -lactalbumin Sepharose, Q-Sepharose, UDP-hexanolamine Sepharose, and GalNAc1-4Gal-Synsorb column chromatography. The purified enzyme was resolved into two bands on reducing gels with apparent molecular weights of 62 kDa and 65 kDa, respectively. GalT-3 activity was also localized in the same regions by activity gel analysis and sucrose-density gradient centrifugation of a detergent-solubilized extract of 19-day-old embryonic chicken brain. Purified GalT-3 exhibited apparentK
mS of 33 µm, 22 µm and 14.4mM with respect to the substrates GM2, UDP-galactose, and MnCl2, respectively. Substrate specificity studies with the purified enzyme and a variety of glycosphingolipids, glycoproteins, and synthetic substrates revealed that the enzyme was highly specific only for the glycosphingolipid acceptors, GM2 and GgOse3Cer (asialo-GM2). Ovine-asialo-agalacto submaxillary mucin inhibited the transfer of galactose to GM2 but did not act as an acceptor in the range of concentrations tested. Polyclonal antibodies raised against purified GalT-3 inhibited GalT-3 activityin vitro and Western-immunoblot analysis of purified GalT-3 showed immunopositive bands at 62 and 65 kDa.Abbreviations CNS
central nervous system
- GM1
monosialotetraosylganglioside, Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer
- GM2
monosialotriaosylganglioside, GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer
- DSS
detergent solubilized supernatant
- ECB
embryonic chicken brain
- TBS
Tris-buffered saline 相似文献
20.