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101.
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103.
Mieke M. E. Huijbers Jenny W. Wu Adrie H. Westphal Willem J. H. van Berkel 《Biotechnology journal》2019,14(5)
Thermus thermophilus proline dehydrogenase ( TtProDH) catalyzes the first step in proline catabolism. The thermostable flavoenzyme consists of a distorted triosephosphate isomerase (TIM) barrel and three N‐terminal helices: αA, αB, and αC. Using maltose‐binding protein (MBP) fused constructs, it has been recently demonstrated that helix αC is crucial for TtProDH catalysis and for tetramerization through positioning of helix α8. Here, the structural features that determine the thermostability of TtProDH are reported. Selective disruption of two ion pairs in the dimerization interface of several MBP‐TtProDH variants result in the formation of monomers. The newly created monomers have improved catalytic properties but their melting temperatures are decreased by more than 20 °C. Sequence comparison suggests that one of the ion‐pairs involved in dimerization is unique for ProDHs from Thermus species. In summary, intermolecular ion‐pairs improve the thermostability of TtProDH and a trade‐off is made between thermostability and catalytic activity. 相似文献
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105.
Evolutionary patterns in the antR-Cor gene in the dwarf dogwood complex (Cornus, Cornaceae) 总被引:1,自引:0,他引:1
The evolutionary pattern of the myc-like anthocyanin regulatory gene antR-Cor was examined in the dwarf dogwood species complex (Cornus Subgenus Arctocrania) that contains two diploid species (C. canadensis and C. suecica), their putative hybrids with intermediate phenotypes, and a tetraploid derivative (C. unalaschkensis). Full-length sequences of this gene (∼4 kb) were sequenced and characterized for 47 dwarf dogwood samples representing all
taxa categories from 43 sites in the Pacific Northwest. Analysis of nucleotide diversity indicated departures from neutral
evolution, due most likely to local population structure. Neighbor-joining and haplotype network analyses show that sequences
from the tetraploid and diploid intermediates are much more strongly diverged from C. suecica than from C. canadensis, and that the intermediate phenotypes may represent an ancestral group to C. canadensis rather than interspecific hybrids. Seven amino acid mutations that are potentially linked to myc-like anthocyanin regulatory
gene function correlate with petal colors differences that characterize the divergence between two diploid species and the
tetraploid species in this complex. The evidence provides a working hypothesis for testing the role of the gene in speciation
and its link to the petal coloration. Sequencing and analysis of additional nuclear genes will be necessary to resolve questions
about the evolution of the dwarf dogwood complex. 相似文献
106.
Considerable effort has been invested in determining traits underlying invasiveness. Yet, identifying a set of traits that
commonly confers invasiveness in a range of species has proven elusive, and almost nothing is known about genetic loci affecting
invasive success. Incorporating genetic model organisms into ecologically relevant studies is one promising avenue to begin
dissecting the genetic underpinnings of invasiveness. Molecular biologists are rapidly characterizing genes mediating developmental
responses to diverse environmental cues, i.e., genes for plasticity, as well as to environmental factors likely to impose
strong selection on invading species, e.g., resistance to herbivores and competitors, coordination of life-history events
with seasonal changes, and physiological tolerance of heat, drought, or cold. Here, we give an overview of molecular genetic
tools increasingly used to characterize the genetic basis of adaptation and that may be used to begin identifying genetic
mechanisms of invasiveness. Given the divergent traits that affect invasiveness, “invasiveness genes” common to many clades
are unlikely, but the combination of developmental genetic advances with further evolutionary studies and modeling may provide
a framework for identifying genes that account for invasiveness in related species. 相似文献
107.
Maines MD Miralem T Lerner-Marmarosh N Shen J Gibbs PE 《The Journal of biological chemistry》2007,282(11):8110-8122
Human biliverdin reductase (hBVR), a dual specificity kinase (Ser/Thr/Tyr) is, as protein kinase C (PKC) betaII, activated by insulin and free radicals (Miralem, T., Hu, Z., Torno, M. D., Lelli, K. M., and Maines, M. D. (2005) J. Biol. Chem. 280, 17084-17092; Lerner-Marmarosh, N., Shen, J., Torno, M. D., Kravets, A., Hu, Z., and Maines, M. D. (2005) Proc. Natl. Acad. Sci. U. S. A. 102, 7109-7114). Here, by using 293A cells co-transfected with pcDNA3-hBVR and PKC betaII plasmids, we report the co-immunoprecipitation of the proteins and co-purification in the glutathione S-transferase (GST) pulldown assay. hBVR and PKC betaII, but not the reductase and PKC zeta, transphosphorylated in assay systems supportive of activity of only one of the kinases. PKC betaII K371R mutant protein ("kinase-dead") was also a substrate for hBVR. The reductase increased the Vmax but not the apparent Km values of PKC betaII for myelin basic protein; activation was independent of phospholipids and extended to the phosphorylation of S2, a PKC-specific substrate. The increase in substrate phosphorylation was blocked by specific inhibitors of conventional PKCs and attenuated by sihBVR. The effect of the latter could be rescued by subsequent overexpression of hBVR. To a large extent, the activation was a function of the hBVR N-terminal chain of valines and intact ATP-binding site and the cysteine-rich C-terminal segment. The cobalt protoporphyrin-activated hBVR phosphorylated a threonine in a peptide corresponding to the Thr500 in the human PKC betaII activation loop. Neither serine nor threonine residues in peptides corresponding to other phosphorylation sites of the PKC betaII nor PKC zeta activation loop-derived peptides were substrates. The phosphorylation of Thr500 was confirmed by immunoblotting of hBVR.PKC betaII immunocomplex. The potential biological relevance of the hBVR activation of PKC betaII was suggested by the finding that in cells transfected with the PKC betaII, hBVR augmented phorbol myristate acetate-mediated c-fos expression, and infection with sihBVR attenuated the response. Also, in cells overexpressing hBVR and PKC betaII, as well as in untransfected cells, upon treatment with phorbol myristate acetate, the PKC translocated to the plasma membrane and co-localized with hBVR. hBVR activation of PKC betaII underscores its potential function in propagation of signals relayed through PKCs. 相似文献
108.
Wordsworth S Buchanan J Regan R Davison V Smith K Dyer S Campbell C Blair E Maher E Taylor J Knight SJ 《Genomic Medicine》2007,1(1-2):35-45
Array based comparative genomic hybridisation (aCGH) is a powerful technique for detecting clinically relevant genome imbalance
and can offer 40 to > 1000 times the resolution of karyotyping. Indeed, idiopathic learning disability (ILD) studies suggest
that a genome-wide aCGH approach makes 10–15% more diagnoses involving genome imbalance than karyotyping. Despite this, aCGH
has yet to be implemented as a routine NHS service. One significant obstacle is the perception that the technology is prohibitively
expensive for most standard NHS clinical cytogenetics laboratories. To address this, we investigated the cost-effectiveness
of aCGH versus standard cytogenetic analysis for diagnosing idiopathic learning disability (ILD) in the NHS. Cost data from
four participating genetics centres were collected and analysed. In a single test comparison, the average cost of aCGH was
£442 and the average cost of karyotyping was £117 with array costs contributing most to the cost difference. This difference
was not a key barrier when the context of follow up diagnostic tests was considered. Indeed, in a hypothetical cohort of 100 ILD
children, aCGH was found to cost less per diagnosis (£3,118) than a karyotyping and multi-telomere FISH approach (£4,957).
We conclude that testing for genomic imbalances in ILD using microarray technology is likely to be cost-effective because
long-term savings can be made regardless of a positive (diagnosis) or negative result. Earlier diagnoses save costs of additional
diagnostic tests. Negative results are cost-effective in minimising follow-up test choice. The use of aCGH in routine clinical
practice warrants serious consideration by healthcare providers.
Copyright statement The Corresponding Author has the right to grant on behalf of all authors and does grant on behalf of all authors, an exclusive
licence (or non exclusive for government employees) on a worldwide basis to the BMJ Publishing Group Ltd, and its Licensees
to permit this article (if accepted) to be published in BMJ editions and any other BMJPGL products and to exploit all subsidiary
rights, as set out in our licence (bmj.com/advice/copyright.shtml).
Authorship The authors included on this paper fulfil the criteria of authorship and no one who fulfils the criteria has been excluded
from authorship. The authors made a substantial contribution to the conception, design, analysis and interpretation of data.
They were involved in drafting the article or revising it critically for important intellectual content and approving the
version to be published.
Contributorship Sarah Wordsworth (Guarantor): Planning, conducting and reporting work, interpretation of data, drafting and revising article.
James Buchanan: Conducting and reporting work, interpretation of data, revising article.
Regina Regan: Completing costing questionnaire, providing protocol details, other costing information, interpretation of data,
information about learning disability and genome imbalance and revising article.
Val Davison: Completing costing questionnaire, providing protocol details, sharing overall laboratory experience and drafting
article.
Kim Smith: Completing costing questionnaire, providing protocol details, drafting article.
Sara Dyer: Completing costing questionnaire and providing protocol details.
Carolyn Campbell: Completing costing questionnaire and providing protocol details.
Edward Blair: Critical appraisal of article for clinical content and revising article.
Eddy Maher: Completing costing questionnaire, providing protocol details, sharing overall laboratory experience and drafting
article.
Jenny Taylor: Planning and facilitating work between centres. Drafting and revising article.
Samantha JL Knight: Completing costing questionnaire, providing protocol details, other costing information, interpretation
of data, providing information about learning disability and genome imbalance, drafting and revising article.
Jenny Taylor and Samantha JL Knight contributed equally to the work presented. 相似文献
109.
Objective: Our aim was to examine whether secular trends in childhood overweight and obesity during five decades could be explained by economic growth. Research Methods and Procedures: Annual measurements of height and weight were available for all children born between 1930 and 1983 attending primary school in the Copenhagen Municipality: 165,389 boys and 163,609 girls from the age of 7 through 13 years. After computerization, we calculated BMI (kg/m2) and estimated the prevalence of overweight and obesity, according to international age‐ and gender‐specific criteria, by year of birth and of measurement, and separately by each age group and gender. Economic growth was indicated by the Gross National Product and the overall consumption per capita, adjusted for inflation. Results: The prevalence of overweight occurred in phases: an increase from 1930 until the 1950s, followed by a plateau period between the 1950s and the 1960s and a steep increase thereafter. This pattern was apparent across all age groups and in both genders. Obesity trends showed a similar phase pattern; the prevalence remained relatively stable from 1930 until the 1940s, increased until the mid‐1950s, followed by a plateau until 1965, and thereafter a second steep increase. Obesity trends were similar among boys across all age groups, although only among girls from 11 to 13 years of age. In both genders, increments were most pronounced in the upper BMI percentiles. After stagnation until 1947, the economic growth indicators showed a steady increase; i.e., after the first increase started in overweight and obesity, whether analyzed by year of birth or year of measurement, there were no indications of phases in the rise thereafter. Discussion: Prevalence of overweight and obesity among Danish children rose in phases, which were not paralleled by trends in economic growth. The macroeconomic growth indicators seem inappropriate as proxies for the environmental exposures that have elicited the obesity epidemic. 相似文献
110.
Peterson RS Fernando G Day L Allen TA Chapleau JD Menjivar J Schlinger BA Lee DW 《Developmental neurobiology》2007,67(14):1867-1878
Estrogens can be neuroprotective following traumatic brain injury. Immediately after trauma to the zebra finch hippocampus, the estrogen-synthetic enzyme aromatase is rapidly upregulated in astrocytes and radial glia around the lesion site. Brain injury also induces high levels of cell proliferation. Estrogens promote neuronal differentiation, migration, and survival naturally in the avian brain. We suspect that glia are a source of estrogens promoting cell proliferation after neural injury. To explore this hypothesis, we examined the spatial and temporal relationship between glial aromatase expression and cell proliferation after neural injury in adult female zebra finches. Birds were ovariectomized and given a blank implant or one filled with estradiol; some birds were also administered an aromatase inhibitor or vehicle. All birds received penetrating injuries to the right hippocampus. Twenty-four hours after lesioning, birds were injected once with BrdU to label mitotically active cells and euthanized 2 h, 24 h, or 7 days later. The brains were processed for double-label BrdU and aromatase immunocytochemistry. Injury-induced glial aromatase expression was unaffected by survival time and aromatase inhibition. BrdU labeling was significantly reduced at 24 h by ovariectomy and by aromatase inhibition; effects were partially reversed by E2 replacement. Irrespective of ovariectomy, the densities of aromatase immunoreactive astrocytes and BrdU-labeled cells at known distances from the lesion site were highly correlated. These data suggest that injury-induced glial aromatization may influence the reorganization of injured tissue by providing a rich estrogenic environment available to influence cellular incorporation. 相似文献