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21.
Acceptor specificity of the human leukocyte alpha3 fucosyltransferase: role of FucT-VII in the generation of selectin ligands 总被引:2,自引:2,他引:0
Britten CJ; van den Eijnden DH; McDowell W; Kelly VA; Witham SJ; Edbrooke MR; Bird MI; de Vries T; Smithers N 《Glycobiology》1998,8(4):321-327
The alpha3 fucosyltransferase, FucT-VII, is one of the key
glycosyltransferases involved in the biosynthesis of the sialyl Lewis X
(sLex) antigen on human leukocytes. The sialyl Lewis X antigen
(NeuAcalpha(2-3)Galbeta(1-4)[Fucalpha(1-3)]GlcNAc-R) is an essential
component of the recruitment of leukocytes to sites of inflammation,
mediating the primary interaction between circulating leukocytes and
activated endothelium. In order to characterize the enzymatic properties of
the leukocyte alpha3 fucosyltransferase FucT-VII, the enzyme has been
expressed in Trichoplusia ni insect cells. The enzyme is capable of
synthesizing both sLexand sialyl-dimeric-Lexstructures in vitro , from
3'-sialyl-lacNAc and VIM-2 structures, respectively, with only low levels
of fucose transfer observed to neutral or 3'-sulfated acceptors. Studies
using fucosylated NeuAcalpha(2-3)-(Galbeta(1- 4)GlcNAc)3-Me acceptors
demonstrate that FucT-VII is able to synthesize both di-fucosylated and
tri-fucosylated structures from mono- fucosylated precursors, but
preferentially fucosylates the distal GlcNAc within a polylactosamine
chain. Furthermore, the rate of fucosylation of the internal GlcNAc
residues is reduced once fucose has been added to the distal GlcNAc. These
results indicate that FucT-VII is capable of generating complex selectin
ligands, in vitro , however the order of fucose addition to the lactosamine
chain affects the rate of selectin ligand synthesis.
相似文献
22.
In spite of the increasing application of DNA fingerprinting to natural
populations and to the genetic identification of humans, explicit methods
for estimation of basic population genetic parameters from DNA
fingerprinting data have not been developed. Contributing to this omission
is the inability to determine, for multilocus fingerprinting probes,
relatively important genetic information, such as the number of loci, the
number of alleles, and the distribution of these alleles into specific
loci. One of the most useful genetic parameters that could be derived from
such data would be the average heterozygosity, which has traditionally been
employed to measure the level of genetic variation within populations and
to compare genetic variation among different loci. We derive here explicit
formulas for both the estimation of average heterozygosity at multiple
hypervariable loci and a maximum value for this estimate. These estimates
are based upon the DNA restriction-pattern matrices that are typical for
fingerprinting studies of humans and natural populations. For several
empirical data sets from our laboratory, estimates of average and maximal
heterozygosity are shown to be relatively close to each other. Furthermore,
variances of these statistics based on simulation studies are relatively
small. These observations, as well as consideration of the effect of
missing alleles and alternate numbers of loci, suggest that the average
heterozygosity can be accurately estimated using phenotypic DNA fingerprint
patterns, because this parameter is relatively insensitive to the lack of
certain genetic information.
相似文献
23.
The breakpoint cluster region protein, BCR, has protein kinase activity that can auto- and trans-phosphorylate serine, threonine and tyrosine residues. BCR has been implicated in chronic myelogenous leukaemia as well as important signalling pathways, and as such its interaction with 14-3-3 is of major interest. 14-3-3tau and zeta isoforms have been shown previously to be phosphorylated in vitro and in vivo by BCR kinase on serine and threonine residue(s) but site(s) were not determined. Phosphorylation of 14-3-3 isoforms at distinct sites is an important mode of regulation that negatively affects interaction with Raf kinase and Bax, and potentially influences the dimerization of 14-3-3. In this study we have further characterized the BCR-14-3-3 interaction and have identified the site phosphorylated by BCR. We show here that BCR interacts with at least five isoforms of 14-3-3 in vivo and phosphorylates 14-3-3tau on Ser233 and to a lesser extent 14-3-3zeta on Thr233. We have previously shown that these two isoforms are also phosphorylated at this site by casein kinase 1, which, in contrast to BCR, preferentially phosphorylates 14-3-3zeta. 相似文献
24.
EMMA L. WRIGHT COLIN R. BLACK ALEXANDER W. CHEESMAN TREVOR DRAGE DAVID LARGE BENJAMIN L. TURNER SOFIE SJÖGERSTEN 《Global Change Biology》2011,17(9):2867-2881
Tropical peatlands play an important role in the global carbon cycling but little is known about factors regulating carbon dioxide (CO2) and methane (CH4) fluxes from these ecosystems. Here, we test the hypotheses that (i) CO2 and CH4 are produced mainly from surface peat and (ii) that the contribution of subsurface peat to net C emissions is governed by substrate availability. To achieve this, in situ and ex situ CO2 and CH4 fluxes were determined throughout the peat profiles under three vegetation types along a nutrient gradient in a tropical ombrotrophic peatland in Panama. The peat was also characterized with respect to its organic composition using 13C solid state cross‐polarization magic‐angle spinning nuclear magnetic resonance spectroscopy. Deep peat contributed substantially to CO2 effluxes both with respect to actual in situ and potential ex situ fluxes. CH4 was produced throughout the peat profile with distinct subsurface peaks, but net emission was limited by oxidation in the surface layers. CO2 and CH4 production were strongly substrate‐limited and a large proportion of the variance in their production (30% and 63%, respectively) was related to the quantity of carbohydrates in the peat. Furthermore, CO2 and CH4 production differed between vegetation types, suggesting that the quality of plant‐derived carbon inputs is an important driver of trace gas production throughout the peat profile. We conclude that the production of both CO2 and CH4 from subsurface peat is a substantial component of the net efflux of these gases, but that gas production through the peat profile is regulated in part by the degree of decomposition of the peat. 相似文献
25.
Chan YW Mohr R Millard AD Holmes AB Larkum AW Whitworth AL Mann NH Scanlan DJ Hess WR Clokie MR 《Molecular biology and evolution》2011,28(8):2269-2274
DNA polymerase γ is a family A DNA polymerase responsible for the replication of mitochondrial DNA in eukaryotes. The origins of DNA polymerase γ have remained elusive because it is not present in any known bacterium, though it has been hypothesized that mitochondria may have inherited the enzyme by phage-mediated nonorthologous displacement. Here, we present an analysis of two full-length homologues of this gene, which were found in the genomes of two bacteriophages, which infect the chlorophyll-d containing cyanobacterium Acaryochloris marina. Phylogenetic analyses of these phage DNA polymerase γ proteins show that they branch deeply within the DNA polymerase γ clade and therefore share a common origin with their eukaryotic homologues. We also found homologues of these phage polymerases in the environmental Community Cyberinfrastructure for Advanced Microbial Ecology Research and Analysis (CAMERA) database, which fell in the same clade. An analysis of the CAMERA assemblies containing the environmental homologues together with the filter fraction metadata indicated some of these assemblies may be of bacterial origin. We also show that the phage-encoded DNA polymerase γ is highly transcribed as the phage genomes are replicated. These findings provide data that may assist in reconstructing the evolution of mitochondria. 相似文献
26.
1. Benthic macroinvertebrates (MI) are commonly used to assess freshwater ecosystems with the reference condition approach. Such assessments necessitate control for natural community variation, either by categorical typologies or by predictive models that have been widely and successfully developed for running water biota but not previously for lake profundal invertebrates. 2. We evaluated four modelling techniques [multivariate regression tree (MRT), limiting environmental differences, nonparametric multiplicative regression (NPMR) and River Invertebrate Prediction And Classification System (RIVPACS) and the operative Finnish lake typology for assessing taxonomic completeness (observed‐to‐expected number of taxa, O/E) of profundal MI assemblages. We used data from 74 and 33 minimally disturbed reference lake basins for calibration and validation of the approaches, respectively, and 72 test basins subject to various anthropogenic pressures to evaluate sensitivity to detect impact. Either all predicted taxa (threshold probability of capture Pt = 0+) or only those predicted to be captured with ≥0.25 probability were used to calculate O/E. 3. With Pt = 0.25, all four modelling approaches were accurate (mean O/E = 0.966–1.053) but imprecise (SD of O/E = 0.279–0.304) in predicting the fauna actually observed in validation sites. All models were subtly more precise than a null model (mean 1.038, SD 0.343) or the typology (1.046, 0.327). The taxon‐specific NPMR model was slightly more precise than the other three models based on site groupings. 4. The O/E values correlated relatively weakly (r = 0.55–0.86) among the approaches, which thus produced contrasting lake‐specific assessments, despite their seemingly comparable performances. Indeed, typology, suggesting that MI assemblages were impaired in 56% of test sites, was more sensitive than the other approaches (26–46%) as an indicator of human‐induced deterioration. However, this greater ostensible sensitivity seemed to be biased, as lake morphometry, a main driver of natural community variation, remained uncontrolled by the typology. 5. Generally, our exercise illustrates the inconclusiveness of the common validation criteria for the assessment methods. The apparent poor predictability of the profundal fauna, irrespective of the method, may partly stem from large observation error, which could be alleviated by more intensive sampling. However, instead of an O/E‐taxa index, some other metric encompassing quantitative aspects might be preferable for assessing these species‐poor communities. 相似文献
27.
28.
Hayley M Bennett Hoi Ping Mok Effrossyni Gkrania-Klotsas Isheng J Tsai Eleanor J Stanley Nagui M Antoun Avril Coghlan Bhavana Harsha Alessandra Traini Diogo M Ribeiro Sascha Steinbiss Sebastian B Lucas Kieren SJ Allinson Stephen J Price Thomas S Santarius Andrew J Carmichael Peter L Chiodini Nancy Holroyd Andrew F Dean Matthew Berriman 《Genome biology》2014,15(11)
29.
To improve recombinant human bone morphogenetic protein-2 (rhBMP-2) yield, cell lines stably expressing hBMP2 were cultured in the presence of polyarginine peptide IND-1 and showed up to 6-fold increase in the yield of mature BMP-2.
Repeated addition of IND-1 to cell cultures consistently improved BMP-2 yields over 53 days without affecting cell growth
and viability. Investigation of its mechanism of action showed that IND-1 inhibited pro-protein convertase (PC) activity when
incubated with cell lysates. However, when intact cells were cultured with IND-1, no change in cellular PC activity was observed.
Furthermore, knockdown of furin (a prototypical member of the PCs) in cells did not affect their BMP-2 yields, suggesting
furin/PC inhibition is unlikely the mechanism by which IND-1 enhances BMP-2 yields. IND-1 as a medium additive thus enhances
BMP-2 production in mammalian cell expression systems. 相似文献
30.
Chien-Hsing Lin Ling-Hui Li Sheng-Feng Ho Tzu-Po Chuang Jer-Yuarn Wu Yuan-Tsong Chen Cathy SJ Fann 《BMC genetics》2008,9(1):1-9