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1.
Minjuan Shen Mingli Lin Mengqi Zhu Wenxin Zhang Danyang Lu Huanhuan Liu Jingjing Deng Kehua Que Xu Zhang 《Biochimica et Biophysica Acta (BBA)/General Subjects》2019,1863(1):167-181
Since their discovery, matrix vesicles (MVs) containing minerals have received considerable attention for their role in the mineralization of bone, dentin and calcified cartilage. Additionally, MVs' association with collagen fibrils, which serve as the scaffold for calcification in the organic matrix, has been repeatedly highlighted. The primary purpose of the present study was to establish a MVs–mimicking model (PEG-S-ACP/micelle) in vitro for studying the exact mechanism of MVs-mediated extra/intra fibrillar mineralization of collagen in vivo. In this study, high-concentration serine was used to stabilize the amorphous calcium phosphate (S-ACP), which was subsequently mixed with polyethylene glycol (PEG) to form PEG-S-ACP nanoparticles. The nanoparticles were loaded in the polysorbate 80 micelle through a micelle self-assembly process in an aqueous environment. This MVs–mimicking model is referred to as the PEG-S-ACP/micelle model. By adjusting the pH and surface tension of the PEG-S-ACP/micelle, two forms of minerals (crystalline mineral nodules and ACP nanoparticles) were released to achieve the extrafibrillar and intrafibrillar mineralization, respectively. This in vitro mineralization process reproduced the mineral nodules mediating in vivo extrafibrillar mineralization and provided key insights into a possible mechanism of biomineralization by which in vivo intrafibrillar mineralization could be induced by ACP nanoparticles released from MVs. Also, the PEG-S-ACP/micelle model provides a promising methodology to prepare mineralized collagen scaffolds for repairing bone defects in bone tissue engineering. 相似文献
2.
Immediate fragmentation of parental DNA by near-ultraviolet irradiation at 313 nm was measured in cultured skin fibroblasts from normal individuals, patients with Xeroderma pigmentosum of complementation group A (XPA) and Xeroderma pigmentosum variants (XPV) by the alkaline elution procedure. For a dose of 2.25 KJm?2 given at Oo fragmentation was comparable in all cell strains. However, fragmentation was strongly increased relative to Oo in XPV but not in normal fibroblasts and the XPA strains when irradiation was carried out at 37o. From our results it appears that a step in the repair of DNA is abnormal in XPV. 相似文献
3.
Intestinal cholesterol absorption is specifically inhibited by the 2-azetidinone cholesterol absorption inhibitor ezetimibe. Photoreactive ezetimibe analogues specifically label a 145-kDa protein in the brush border membrane of enterocytes from rabbit small intestine identified as aminopeptidase N (CD13). In zebrafish and mouse small intestinal cytosol, a heterocomplex of Mr 52 kDa between annexin II and caveolin 1 was suggested as a target of ezetimibe. In contrast, in the cytosol and brush border membrane vesicles (BBMV) from rabbit small intestine of control animals or rabbits treated with the nonabsorbable cholesterol absorption inhibitor AVE 5530, both annexin II and caveolin 1 were exclusively present as monomers without any heterocomplex formation. Upon immunoprecipitation with annexin II a 52-kDa band was observed after immunostaining with annexin II antibodies, whereas no staining of a 52-kDa band occurred with anti-caveolin 1 antibodies. Vice versa, a 52-kDa band obtained by immunoprecipitation with caveolin 1 antibodies did not stain with annexin II-antibodies. The intensity of the 52-kDa band was dependent on the amount of antibody and was also observed with anti-actin or anti-APN antibodies suggesting that the 52-kDa band is a biochemical artefact. After incubation of cytosol or BBMV with radioactively labelled ezetimibe analogues, no significant amounts of the ezetimibe analogues could be detected in the immunoprecipitate with caveolin-1 or annexin II antibodies. Photoaffinity labelling of rabbit small intestinal BBMV with ezetimibe analogues did not result in labelling of proteins being immunoreactive with annexin II, caveolin 1 or a 52-kDa heterocomplex. These findings indicate that the rabbit small intestine does not contain an annexin II/caveolin 1 heterocomplex as a target for ezetimibe. 相似文献
4.
5.
M.J. Tisdale 《Chemico-biological interactions》1981,34(1):75-83
The tumour growth inhibitor L-2-amino-4-methoxy-trans-3-butenoic acid (Ro07-7957) inhibits serine hydroxymethyltransferase in cytosolic extracts of Walker carcinoma non-competitively with respect to L-serine with an apparent inhibition constant similar to the Km-value for L-serine. The kinetics of inactivation suggest that it reacts as an irreversible substrate analogue. Incubation of Walker cells with Ro07-7957 causes an increase in serine hydroxymethyltransferase activity which is most pronounced at concentration ≤LD50. This increase in enzyme activity does not occur in the presence of cycloheximide. These results suggest that inhibition of serine hydroxymethyltransferase in intact cells is accompanied by an increase in enzyme biosynthesis and that the growth inhibitory property of Ro07-7957 does not involve interference with the conversion of serine to glycine. 相似文献
6.
7.
B. C. Loughman 《Plant and Soil》1987,99(1):63-74
Summary Little is known about the biochemical basis of the genotypic differences in the capacity for ion absorption and transport
shown by many crop species. If these differences reflect the abundance of a specific membrane component or the activity of
an enzyme we need to have some indication of thein vivo operation of these systems in whole plants. Thein vivo assessment of glycolytic enzymes is illustrated by the effects of mannose on the transport of phosphate in maize varieties.
The application of high resolution31P-NMR to the study of intermediary metabolismin vivo is also helpful in following transport capacity.
The five-fold rise in respiratory rate that occurs when freshly cut potato slices are maintained in aerated water for 24 hours
is accompained by the turning on of a wide range of biochemical systems. Major increases in the capacity for absorption of
phosphate from low concentrations (0.1 μM–10 μM) and in the phosphorylative ability of the tissue are seen, indicating the synthesis of a carrier involved in phosphate transport.
These capacities differ markedly between individual tissues of the tuber,i. e. pith, parenchyma, cortex and buds and large differences have been observed between comparable tissue from different varieties.
Varieties grown under similar conditions have been compared and shown to exhibit different kinetics with respect to the development
of the low concentration absorption site and in their sensitivity to the effects of uncouplers such as 2,4-dinitrophenol. 相似文献
8.
Norma J. Greenfield Mussadeq Hussain John Lenard 《Biochimica et Biophysica Acta (BBA)/General Subjects》1987,926(3)
The vacuoles of logarithmic and stationary stage cells were compared by 31P-NMR with regard to pH, orthophosphate (Pi) content and average size of polyphosphate. The vacuoles of stationary cells had lower pH higher Pi content, and polyphosphates of longer average chain lenght, although total polyphosphate content was about the same as in logarithmic cells. The lower vacuolar pH in stationary cells was the major cause of a larger cytoplasmic-vacuolar pH gradient. Addition of NH4Cl, (NH4)2SO4, methylamine or amantadine at pH 8 to cells in either stage caused an icnrease in both cytoplasmic and vacuolar pH, with little or no change in the cytoplasmic-vacuolar pH gradient. However, the administration of ammonium salts to the cells at pH 8.0 resulted in rapid hydrolysis of the intravacuolar polyphosphate to tripolyphosphate and Pi, with attendant redistribution of Pi between the vacuolar and cytoplasmic compartments. 相似文献
9.
Summary Time-course experiments of phosphate uptake by size-fractionated phytoplankton were conducted in oligotrophic Kennedy and Sproat Lakes. The objective was to determine if large phytoplankton obtained more phosphate than smaller cells, when the nutrient was present at higher concentrations. Studies at Kennedy Lake revealed that uptake rates in the 0.2–3.0 m fraction were very sensitive to the time they were exposed to elevated concentrations; rates determined over the 60–120 min interval were less than 30% of those recorded over the 0–60 min interval. In contrast, there was little difference in uptake rates over these intervals for cells>3.0 m. At Sproat Lake phosphate incorporation into the two size fractions was followed after the aerial fertilization of the lake with inorganic nutrients. Following nutrient addition the proportion of phosphate entering the>3.0 m size fraction increased from ca. 35% to ca. 85%. Despite these observations, it is doubtful that larger cells are able to sequester enough phosphate from pulses to realize the same specific growth rates as their smaller counterparts. 相似文献
10.
The aim of this work was to examine the effect of abrupt changes in temperature in the range 5 to 30°C upon the rate of photosynthetic carbon assimilation in leaves of barley (Hordeum vulgare L.). Measurement of the CO2-assimilation rate in relation to the intercellular partial pressure of CO2 at different temperatures and O2 concentrations and at saturating irradiance showed that as the temperature was decreased photosynthesis was saturated at progressively lower CO2 partial pressures and that the transition between the CO2-limited and ribulose-1,5-bisphosphate-regeneration-limited rate became more abrupt. Feeding of orthophosphate to leaves resulted in an increased rate of CO2 assimilation at lower temperatures at around ambient or higher CO2 partial pressures both in 20% O2 and in 2% O2 and it removed the abruptness in the transition between the CO2-limited and ribulose-1,5-bisphosphate-regeneration-limited rates. Phosphate feeding tended to inhibit carbon assimilation at higher temperatures. The response of carbon assimilation to temperature was altered by feeding orthophosphate, by changing the concentrations of CO2 or of O2 or by leaving plants in the dark at 4°C for several hours. Similarly, the response of carbon assimilation to phosphate feeding or to changes in 2% O2 was altered by leaving the plants in the dark at 4°C. The mechanism of limitation of photosynthesis by an abrupt lowering of temperature is discussed in the light of the results.Abbreviations A
rate of CO2 assimilation
-
P
i
intercellular partial pressure of CO2
- RuBP
ribulose-1,5-bisphosphate 相似文献