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81.
Previous investigations on the monkey kidney COS cell line demonstrated the
weak expression of fucosylated cell surface antigens and presence of
endogenous fucosyltransferase activities in cell extracts. RT-PCR analyses
have now revealed expression of five homologs of human fucosyltransferase
genes, FUT1, FUT4, FUT5, FUT7, and FUT8, in COS cell mRNA. The enzyme in
COS cell extracts acting on unsialylated Type 2 structures is closely
similar in its properties to the alpha1,3- fucosyltransferase encoded by
human FUT4 gene and does not resemble the product of the FUT5 gene.
Although FUT1 is expressed in the COS cell mRNA, it has not been possible
to demonstrate alpha1,2- fucosyltransferase activity in cell extracts but
the presence of Le(y) and blood-group A antigenic determinants on the cell
surface imply the formation of H-precursor structures at some stage. The
most strongly expressed fucosyltransferase in the COS cells is the
alpha1,6-enzyme transferring fucose to the innermost N -acetylglucosamine
unit in N - glycan chains; this enzyme is similar in its properties to the
product of the human FUT8 gene. The enzymes resembling the human FUT4 and
FUT8 gene products both had pH optima of 7.0 and were resistant to 10 mM
NEM. The incorporation of fucose into asialo-fetuin was optimal at 5.5 and
was inhibited by 10 mM NEM. This result initially suggested the presence of
a third fucosyltransferase expressed in the COS cells but we have now shown
that triantennary N- glycans with terminal nonreducing galactose units,
similar to those present in asialo-fetuin, are modified by a weak
endogenous beta-galactosidase in the COS cell extracts and thereby rendered
suitable substrates for the alpha1,6- fucosyltransferase.
相似文献
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Rapid, sequential changes in surface morphology of PC12 pheochromocytoma cells in response to nerve growth factor 总被引:20,自引:17,他引:20 下载免费PDF全文
The effect of nerve growth factor (NGF), a substance that promotes the differentiation and maintenance of certain neurons, was studied via scanning electron microscopy utilizing the PC12 clonal NGF-responsive pheochromocytoma cell line. After 2-4 d of exposure to NGF, these cells acquire many of the properties of normal sympathic neurons. However, by phase microscopy, no changes are discernible within the first 12-18 h. Since the primary NGF receptor appears to be a membrane receptor, it seemed likely that some of the initial responses to the factor may be surface related. PC12 cells maintained without NGF are round to ovoid and have numerous microvilli and small blebs. After the addition of NGF, there is a rapidly initiated sequential change in the cell surface. Ruffles appear over the dorsal surface of the cells with 1 min, become prominent by 3 min, and almost disappear by 7 min. Microvilli, conversely, disappear as the dorsal ruffles become prominent. Ruffles are seen at the the periphery of cell at 3 min, are prominent on most of the cells by 7 min and are gone by 15 min. The surface remains smooth from 15 min until 45 min when large blebs appear. The large blebs are present on most cells at 2 h and are gone by 4 h. The surface remains relatively smooth until 6-7 h of NGF treatment, when microvilli reappear as small knobs. These microvilli increase in both number and length to cover the cell surface by 10 h. These changes were not observed with other basic proteins, with α-bungarotoxin (which binds specifically to PC12 membranes), and were not affected by an RNA synthesis inhibitor that blocks initiation of neurite outgrowth. Changes in the cell surface architecture appear to be among the earlist NGF responses yet detected and may represent or reflect primary events in the mechanism of the factor’s action. 相似文献
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Alan D. Steinman Mary E. Ogdahl Kelly Wessell Bopi Biddanda Scott Kendall Stephen Nold 《Aquatic Ecology》2011,45(4):439-454
We investigated functional and structural responses of periphyton communities to simulated nonpoint source (NPS) pollution over a 2-year period. Periphyton communities were examined in three unique but hydrologically connected habitats in the lower Muskegon River: river, wetland, and lake. Our study addressed several major questions: (1) what are the effects of nonpoint source pollutants on periphyton structure and function, (2) to what degree do environmental conditions, including the pollutant delivery system and hydrologic conditions, affect periphyton response, and (3) do periphyton communities show synchronous responses to nonpoint source pollution? Nutrients were added over a 21-day period in each of the four seasons over 2 years. The effect of road salt on metabolism was evaluated in the winter and spring seasons in the river habitat only. In general, the periphyton community showed complex responses to the NPS amendments, but there was little evidence of nutrient limitation in this system based on community structure, biomass accrual, metabolism, or alkaline phosphatase activity. This result was unexpected given the relatively low ambient concentrations of phosphorus. We discuss the possibilities that nutrient delivery was ineffective or that herbivory and/or irradiance might have constrained periphyton growth, but ultimately conclude that our results suggest that periphyton communities in the lower Muskegon River watershed were (1) nutrient-replete, (2) tolerant of short-term salt additions, and (3) influenced more by local conditions (habitat) than by regional (watershed) factors. In systems where nonpoint source pollutant delivery is diffuse and hydrologic residence time is short, the influence of nonpoint source pollutants on periphyton may be either modest or too difficult to detect using traditional endpoints. 相似文献
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Alan D. Steinman Mary E. Ogdahl Maggie Weinert Donald G. Uzarski 《Aquatic Ecology》2014,48(2):143-159
This study examined the influence of water-level fluctuation (WLF) on sediment–water nutrient exchange in the Laurentian Great Lakes. Water levels in the Laurentian Great Lakes have been below the long-term mean for the past 15 years, causing the exposure of sediments that previously have been either continuously inundated or periodically exposed. The magnitude, duration, and frequency of WLF, as well as land-use history, each can influence the amount and type of sediment–water nutrient exchange. We collected sediment cores from relatively pristine coastal wetlands located on Beaver and Garden Islands in northern Lake Michigan. Sediment cores were taken from different water depths to simulate WLF magnitude; desiccation time was experimentally manipulated to simulate WLF duration. At these relatively pristine wetlands, desiccation time and water depth significantly influenced flux. However, nutrient exchange did not behave in a consistent fashion; phosphorus, nitrate, ammonium, and sulfate flux varied based on sediment exposure history and desiccation time. Sediment–water nutrient exchange rates also were compared to prior measurements taken from more impacted coastal wetlands in southern Lake Michigan and Saginaw Bay in Lake Huron. This comparison revealed a stronger influence of anthropogenic stress than desiccation time, with impacted wetland sediments releasing more soluble reactive phosphorus, sulfate, and ammonium, and retaining more nitrate, than pristine wetlands. Our results indicate that WLFs have the potential to influence sediment–water nutrient exchange, which may influence system productivity, but environmental context can override this influence. 相似文献
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Anke Wesselius Martijn JL Bours Niklas R Jørgensen James Wiley Ben Gu Svenjhalmar van Helden Lodewijk van Rhijn Pieter C Dagnelie 《Purinergic signalling》2013,9(1):123-130
In the present study we investigated whether single nucleotide polymorphisms (SNPs) in the P2RX4, which alter the P2X4R function, are associated with the development of osteoporosis and whether an interaction between the P2X4R and P2X7R confer a synergistic effect of these two receptors on osteoporosis risk. Patients with fracture (690 females and 231 males, aged ≥50 years) were genotyped for three non-synonymous P2X4R SNPs. Bone mineral density (BMD) was measured at the total hip, lumbar spine, and femoral neck. Subject carrying the variant allele of the Tyr315Cys polymorphism showed a 2.68-fold (95 % CI, 1.20–6.02) higher risk of osteoporosis compared with wild-type subject. Furthermore, significant lower lumbar spine BMD values were observed in subjects carrying the Cys315 allele as compared with wild-type (0.85 ± 0.17 and 0.93 ± 0.17 g/cm2, respectively; p < 0.001). Assuming a recessive model, carriers of the variant allele of the Ser242Gly polymorphism showed increased BMD values at the lumbar spine compare to wild-type subject (1.11 ± 0.35 and 0.92 ± 0.17 g/cm2, respectively; p = 0.0045). This is the first study demonstrating an association of non-synonymous polymorphisms in the P2RX4 and the risk of osteoporosis, suggesting a role of the P2X4R in the regulation of bone mass.