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1.
Biomater Sci ; 7(5): 1801-1804, 2019 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-30869657

RESUMO

Here we report a novel aspect of molecular chaperone prefoldin (PFD) as a biomaterial in the biocatalytic synthesis of gold nanoparticles (AuNPs) using glycerol dehydrogenase (GLD). We found that PFD could inhibit the aggregation of AuNPs during the biosynthesis, leading to the formation of AuNPs with controlled size distribution.


Assuntos
Ouro/química , Ouro/metabolismo , Nanopartículas Metálicas , Chaperonas Moleculares/metabolismo , Tamanho da Partícula , Biocatálise , Pyrococcus horikoshii/metabolismo , Desidrogenase do Álcool de Açúcar/metabolismo
2.
Front Plant Sci ; 10: 1809, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-32082354

RESUMO

Pubescence color of soybean is controlled by two genes, T and Td. In the presence of a dominant T allele, dominant and recessive alleles of the Td locus generate tawny and light tawny (or near-gray) pubescence, respectively. Flavones, responsible for pubescence color, are synthesized via two copies of flavone synthase II genes (FNS II-1 and FNS II-2). This study was conducted to map and clone the Td gene. Genetic and linkage analysis using an F2 population and F3 families derived from a cross between a Clark near-isogenic line with light tawny pubescence (genotype: TT tdtd) and a Harosoy near-isogenic line with tawny pubescence (TT TdTd) revealed a single gene for pubescence color around the end of chromosome 3. Genome sequence alignment of plant introductions revealed an association between premature stop codons in Glyma.03G258700 (R2R3 MYB transcription factor) and recessive td allele. Cultivars and lines having near-gray or light tawny pubescence and a gray pubescence cultivar with td allele had premature stop codons in the gene. These results suggest that Glyma.03G258700 corresponds to the Td gene. It was predominantly expressed in pubescence. Compared to a tawny pubescence line, a near-isogenic line with td allele produced extremely small amounts of transcripts of Glyma.03G258700, FNS II-1, and FNS II-2 in pubescence. The promoter of FNS II-1 and FNS II-2 shared cis-acting regulatory elements for binding of MYB proteins. These results suggest that the wild type of Glyma.03G258700 protein may bind to the promoter of FNS II genes and upregulate their expression, resulting in increased flavone content and deeper pubescence color. In contrast, mutated Glyma.03G258700 protein may fail to upregulate the expression of FNS II genes, resulting in decreased flavone content and dilute pubescence color.

3.
Steroids ; 123: 43-49, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28502858

RESUMO

BACKGROUND: Urine has been utilized as a source of biomarkers in renal disease. However, urinary lipids have not attracted much attention so far. Here we studied urinary cholesteryl ester (CE) and its relevance in renal disease. METHODS: Quantitative analysis of CE molecular species in serum, urinary supernatant, and urinary sediment from patients with renal disease (N=64) and non-renal disease (N=23) was carried out using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and deuterated CEs as internal standards. RESULTS: Validation study showed good precision and accuracy of LC-MS/MS. Many CE species were detected in the urinary sediment and supernatant in the renal disease group, whereas only a few CE species were detected in the other group. In the renal disease group, the sum of the concentrations of all CE species showed a significant correlation between the sediment and the supernatant from urinary samples (r=0.876, p<0.001); however, the composition of CEs was significantly different between them. Further, the composition of CEs of the supernatant was similar to that of the serum. CONCLUSIONS: Our LC-MS/MS analysis uncovered a distinct CE profile in urinary sediment from patients with renal disease, suggesting a possible contribution of CEs in urothelial cells to the development of renal disease.


Assuntos
Ésteres do Colesterol/urina , Nefropatias/diagnóstico , Nefropatias/urina , Calibragem , Ésteres do Colesterol/sangue , Cromatografia Líquida , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Espectrometria de Massas em Tandem
4.
Phys Chem Chem Phys ; 19(15): 9843-9848, 2017 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-28352875

RESUMO

The formation process and atomic structures were investigated for single pyrazine molecular junctions sandwiched by three different Au, Ag, and Cu electrodes using a mechanically controllable break junction technique in ultrahigh vacuum conditions at 300 K. We demonstrated that the formation process of the single-molecule junction crucially depended on the choice of the metal electrodes. While single-molecule junction showing two distinct conductance states were found for the Au electrodes, only the single conductance state was evident for the Ag electrodes, and there was no junction formation for the Cu electrodes. These results suggested that metal-molecule interaction dominates the formation process and probability of the single-molecule junction. In addition to the metal-molecule interaction, temperature affected the formation process of the single-molecule junction. The single pyrazine molecular junction formed between Au electrodes exhibited significant temperature dependence where the junction-formation probability was about 8% at 300 K, while there was no junction-formation at 100 K. Instead of the junction formation, an Au atomic wire was formed at the low temperature. This study provides insight into the tuning of the junction-forming process for single-molecule junctions, which is needed to construct device structures on a single molecule scale.

5.
Theor Appl Genet ; 130(4): 743-755, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28097398

RESUMO

KEY MESSAGE: Greatest potential, QTLs for hypoxia and waterlogging tolerance in soybean roots were detected using a new phenotypic evaluation method. Waterlogging is a major environmental stress limiting soybean yield in wet parts of the world. Root development is an important indicator of hypoxia tolerance in soybean. However, little is known about the genetic control of root development under hypoxia. This study was conducted to identify quantitative trait loci (QTLs) responsible for root development under hypoxia. Recombinant inbred lines (RILs) developed from a cross between a hypoxia-sensitive cultivar, Tachinagaha, and a tolerant landrace, Iyodaizu, were used. Seedlings were subjected to hypoxia, and root development was evaluated with the value change in root traits between after and before treatments. We found 230 polymorphic markers spanning 2519.2 cM distributed on all 20 chromosomes (Chrs.). Using these, we found 11 QTLs for root length (RL), root length development (RLD), root surface area (RSA), root surface area development (RSAD), root diameter (RD), and change in average root diameter (CARD) on Chrs. 11, 12, 13 and 14, and 7 QTLs for hypoxia tolerance of these root traits. These included QTLs for RLD and RSAD between markers Satt052 and Satt302 on Chr. 12, which are important markers of hypoxia tolerance in soybean; those QTLs were stable between 2 years. To validate the QTLs, we developed a near-isogenic line with the QTL region derived from Iyodaizu. The line performed well under both hypoxia and waterlogging, suggesting that the region contains one or more genes with large effects on root development. These findings may be useful for fine mapping and positional cloning of gene responsible for root development under hypoxia.


Assuntos
Glycine max/genética , Hipóxia/genética , Raízes de Plantas/crescimento & desenvolvimento , Locos de Características Quantitativas , Estresse Fisiológico , Mapeamento Cromossômico , Ligação Genética , Marcadores Genéticos , Raízes de Plantas/genética , Glycine max/crescimento & desenvolvimento
6.
Plant Mol Biol ; 92(4-5): 445-456, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27561783

RESUMO

KEY MESSAGE: Flavonoids are important secondary metabolites in plants. Sugar-sugar glycosyltransferases are involved in the final step of flavonoid biosynthesis and contribute to the structural diversity of flavonoids. This manuscript describes the first cloning of a sugar-sugar glucosyltransferase gene in the UGT family that attaches glucose to the 6″-position of sugar bound to a flavonol. The results provide a glimpse on the possible evolution of sugar-sugar glycosyltransferase genes and identify putative amino acids responsible for the recognition of the hydroxyl group of the sugar moiety and specification of sugar. A scheme for the genetic control of flavonol glycoside biosynthesis is proposed. Flavonol glycosides (FGs) are predominant in soybean leaves and they show substantial differences among genotypes. In previous studies, we identified two flavonoid glycoside glycosyltransferase genes that segregated in recombinant inbred lines developed from a cross between cultivars Nezumisaya and Harosoy; one was responsible for the attachment of glucose to the 2″-position of glucose or galactose that is bound to the 3-position of kaempferol and the other was involved in the attachment of glucose to the 6″-position. This study was conducted to clone and characterize the 6″-glucosyltransferase gene. Linkage mapping indicated that the gene was located in the molecular linkage group I (chromosome 20). Based on the genome sequence, we cloned a candidate cDNA, GmF3G6"Gt from Harosoy but the corresponding cDNA could not be amplified by PCR from Nezumisaya. The coding region of GmF3G6″Gt in Harosoy is 1386 bp long encoding 462 amino acids. This gene was not expressed in leaves of Nezumisaya. The GmF3G6″Gt recombinant protein converted UDP-glucose and kaempferol 3-O-glucoside or kaempferol 3-O-galactoside to kaempferol 3-O-glucosyl-(1→6)-glucoside or kaempferol 3-O-glucosyl-(1→6)-galactoside, respectively. These results indicate that GmF3G6″Gt encodes a flavonol 3-O-glucoside/galactoside (1→6) glucosyltransferase and corresponds to the Fg1 gene. GmF3G6″Gt had an amino acid similarity of 82 % with GmF3G6″Rt encoding flavonol 3-O-glucoside/galactoside (1→6) rhamnosyltransferase, suggesting a recent evolutionary divergence of the two genes. This may be the first cloning of a sugar-sugar glucosyltransferase gene in the UGT family that attaches glucose to the 6″-position of sugar bound to a flavonol. A scheme for the control of FG biosynthesis is proposed.


Assuntos
Glucosiltransferases/genética , Glucosiltransferases/metabolismo , Glycine max/enzimologia , Glycine max/genética , Mapeamento Cromossômico , Clonagem Molecular , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
7.
Breed Sci ; 66(3): 407-15, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27436951

RESUMO

Time to flowering and maturity in soybean is controlled by loci E1 to E5, and E7 to E9. These loci were assigned to molecular linkage groups (MLGs) except for E5. This study was conducted to map the E5 locus using F2 populations expected to segregate for E5. F2 populations were subjected to quantitative trait locus (QTL) analysis for days to flowering (DF) and maturity (DM). In Harosoy-E5 × Clark-e2 population, QTLs for DF and DM were found at a similar position with E2. In Harosoy × Clark-e2E5 population, QTLs for DF and DM were found in MLG D1a and B1, respectively. In Harosoy-E5Dt2 × Clark-e2 population, a QTL for DF was found in MLG B1. Thus, results from these populations were not fully consistent, and no candidate QTL for E5 was found. In Harosoy × PI 80837 population, from which E5 was originally identified, QTLs corresponding to E1 and E3 were found, but none for E5 existed. Harosoy and PI 80837 had the e2-ns allele whereas Harosoy-E5 had the E2-dl allele. The E2-dl allele of Harosoy-E5 may have been generated by outcrossing and may be responsible for the lateness of Harosoy-E5. We conclude that a unique E5 gene may not exist.

8.
Nanotechnology ; 27(29): 295203, 2016 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-27291763

RESUMO

A highly stable experimental setup was developed for the measurement of shot noise in atomic contacts and molecular junctions to determine the number of atoms or molecules present. The use of a nano-fabricated mechanically controllable break junction (MCBJ) electrode improved the overall stability of the experimental setup. The improved stability of the system and optimization of measurement system enabled us to comprehensively investigate the shot noise as well as charge transport properties in Au atomic contacts and molecular junctions. We present a solid proof that the number of atoms (cross sectional atom) in the Au atomic contacts was exactly one. In the atomic contacts, contribution from the additional channels was under the detection limit. Furthermore, the effect of molecular adsorption on the charge transport in the Au atomic contact was investigated. Additional transport channels were opened by exposing pyrazine molecules to the Au contacts, which gave rise to an increase in the Fano factor in the shot noise.

9.
Structure ; 23(12): 2280-2290, 2015 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-26602186

RESUMO

Steroid receptor drugs have been available for more than half a century, but details of the ligand binding mechanism have remained elusive. We solved X-ray structures of the glucocorticoid and mineralocorticoid receptors to identify a conserved plasticity at the helix 6-7 region that extends the ligand binding pocket toward the receptor surface. Since none of the endogenous ligands exploit this region, we hypothesized that it constitutes an integral part of the binding event. Extensive all-atom unbiased ligand exit and entrance simulations corroborate a ligand binding pathway that gives the observed structural plasticity a key functional role. Kinetic measurements reveal that the receptor residence time correlates with structural rearrangements observed in both structures and simulations. Ultimately, our findings reveal why nature has conserved the capacity to open up this region, and highlight how differences in the details of the ligand entry process result in differential evolutionary constraints across the steroid receptors.


Assuntos
Sequência Conservada , Receptores de Glucocorticoides/química , Receptores de Mineralocorticoides/química , Sequência de Aminoácidos , Sítios de Ligação , Evolução Molecular , Humanos , Dados de Sequência Molecular , Ligação Proteica , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/metabolismo , Receptores de Mineralocorticoides/genética , Receptores de Mineralocorticoides/metabolismo
10.
Genome ; 58(8): 365-74, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26360633

RESUMO

The R gene of soybean, presumably encoding a MYB transcription factor, controls seed coat color. The gene consists of multiple alleles, R (black), r-m (black spots and (or) concentric streaks on brown seed), and r (brown seed). This study was conducted to determine the structure of the MYB transcription factor gene in a near-isogenic line (NIL) having r-m allele. PCR amplification of a fragment of the candidate gene Glyma.09G235100 generated a fragment of about 1 kb in the soybean cultivar Clark, whereas a fragment of about 14 kb in addition to fragments of 1 and 1.4 kb were produced in L72-2040, a Clark 63 NIL with the r-m allele. Clark 63 is a NIL of Clark with the rxp and Rps1 alleles. A DNA fragment of 13 060 bp was inserted in the intron of Glyma.09G235100 in L72-2040. The fragment had the CACTA motif at both ends, imperfect terminal inverted repeats (TIR), inverse repetition of short sequence motifs close to the 5' and 3' ends, and a duplication of three nucleotides at the site of integration, indicating that it belongs to a CACTA-superfamily transposable element. We designated the element as Tgm11. Overall nucleotide sequence, motifs of TIR, and subterminal repeats were similar to those of Tgm1 and Tgs1, suggesting that these elements comprise a family.


Assuntos
Elementos de DNA Transponíveis/genética , Glycine max/genética , Sementes/genética , Proteínas de Soja/genética , Fatores de Transcrição/genética , Alelos , Sequência de Aminoácidos , Sequência de Bases , Genes de Plantas , Loci Gênicos , Íntrons , Dados de Sequência Molecular , Mutação , Reação em Cadeia da Polimerase , Polimorfismo Genético , Sementes/metabolismo , Alinhamento de Sequência , Proteínas de Soja/biossíntese , Glycine max/metabolismo , Sequências Repetidas Terminais , Fatores de Transcrição/metabolismo
11.
PLoS One ; 10(6): e0128527, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26039079

RESUMO

Physical dormancy, a structural feature of the seed coat known as hard seededness, is an important characteristic for adaptation of plants against unstable and unpredictable environments. To dissect the molecular basis of qHS1, a quantitative trait locus for hard seededness in soybean (Glycine max (L) Merr.), we developed a near-isogenic line (NIL) of a permeable (soft-seeded) cultivar, Tachinagaha, containing a hard-seed allele from wild soybean (G. soja) introduced by successive backcrossings. The hard-seed allele made the seed coat of Tachinagaha more rigid by increasing the amount of ß-1,4-glucans in the outer layer of palisade cells of the seed coat on the dorsal side of seeds, known to be a point of entrance of water. Fine-mapping and subsequent expression and sequencing analyses revealed that qHS1 encodes an endo-1,4-ß-glucanase. A single-nucleotide polymorphism (SNP) introduced an amino acid substitution in a substrate-binding cleft of the enzyme, possibly reducing or eliminating its affinity for substrates in permeable cultivars. Introduction of the genomic region of qHS1 from the impermeable (hard-seeded) NIL into the permeable cultivar Kariyutaka resulted in accumulation of ß-1,4-glucan in the outer layer of palisade cells and production of hard seeds. The SNP allele found in the NIL was further associated with the occurrence of hard seeds in soybean cultivars of various origins. The findings of this and previous studies may indicate that qHS1 is involved in the accumulation of ß-1,4-glucan derivatives such as xyloglucan and/or ß-(1,3)(1,4)-glucan that reinforce the impermeability of seed coats in soybean.


Assuntos
Celulase/genética , Glycine max/genética , Proteínas de Plantas/genética , Polimorfismo de Nucleotídeo Único , Sementes/genética , Alelos , Sequência de Aminoácidos , Substituição de Aminoácidos , Transporte Biológico , Celulase/metabolismo , Cruzamentos Genéticos , Expressão Gênica , Glucanos/metabolismo , Dados de Sequência Molecular , Permeabilidade , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Locos de Características Quantitativas , Sementes/metabolismo , Alinhamento de Sequência , Glycine max/metabolismo , Água/metabolismo
12.
BMC Plant Biol ; 15: 126, 2015 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-26002063

RESUMO

BACKGROUND: Flavonol glycosides (FGs) are major components of soybean leaves and there are substantial differences in FG composition among genotypes. The first objective of this study was to identify genes responsible for FG biosynthesis and to locate them in the soybean genome. The second objective was to clone the candidate genes and to verify their function. Recombinant inbred lines (RILs) were developed from a cross between cultivars Nezumisaya and Harosoy. RESULTS: HPLC comparison with authentic samples suggested that FGs having glucose at the 2″-position of glucose or galactose that is bound to the 3-position of kaempferol were present in Nezumisaya, whereas FGs of Harosoy were devoid of 2″-glucose. Conversely, FGs having glucose at the 6″-position of glucose or galactose that is bound to the 3-position of kaempferol were present in Harosoy, whereas these FGs were absent in Nezumisaya. Genetic analysis suggested that two genes control the pattern of attachment of these sugar moieties in FGs. One of the genes may be responsible for attachment of glucose to the 2″-position, probably encoding for a flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase. Nezumisaya may have a dominant whereas Harosoy may have a recessive allele of the gene. Based on SSR analysis, linkage mapping and genome database survey, we cloned a candidate gene designated as GmF3G2″Gt in the molecular linkage group C2 (chromosome 6). The open reading frame of GmF3G2″Gt is 1380 bp long encoding 459 amino acids with four amino acid substitutions among the cultivars. The GmF3G2″Gt recombinant protein converted kaempferol 3-O-glucoside to kaempferol 3-O-sophoroside. GmF3G2″Gt of Nezumisaya showed a broad activity for kaempferol/quercetin 3-O-glucoside/galactoside derivatives but it did not glucosylate kaempferol 3-O-rhamnosyl-(1 → 4)-[rhamnosyl-(1 → 6)-glucoside] and 3-O-rhamnosyl-(1 → 4)-[glucosyl-(1 → 6)-glucoside]. CONCLUSION: GmF3G2″Gt encodes a flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase and corresponds to the Fg3 gene. GmF3G2″Gt was designated as UGT79B30 by the UGT Nomenclature Committee. Based on substrate specificity of GmF3G2″Gt, 2″-glucosylation of flavonol 3-O-glycoside may be irreconcilable with 4″-glycosylation in soybean leaves.


Assuntos
Mapeamento Cromossômico/métodos , Clonagem Molecular/métodos , Flavonóis/metabolismo , Genes de Plantas , Glucosiltransferases/genética , Glycine max/genética , Glicosídeos/metabolismo , Sequência de Aminoácidos , Cromatografia Líquida de Alta Pressão , Flavonóis/análise , Flavonóis/química , Regulação da Expressão Gênica de Plantas , Glucosiltransferases/química , Glucosiltransferases/metabolismo , Glicosídeos/análise , Glicosídeos/química , Endogamia , Padrões de Herança/genética , Dados de Sequência Molecular , Filogenia , Polimorfismo Genético , Recombinação Genética/genética , Especificidade por Substrato
13.
Inorg Chem ; 53(21): 11794-801, 2014 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-25334034

RESUMO

A novel quadruple perovskite oxide CeCu3Fe4O12 has been synthesized under high-pressure and high-temperature conditions of 15 GPa and 1473 K. (57)Fe Mössbauer spectroscopy displays a charge disproportionation transition of 4Fe(3.5+) → 3Fe(3+) + Fe(5+) below ∼270 K, whereas hard X-ray photoemission and soft X-ray absorption spectroscopy measurements confirm that the Ce and Cu valences are retained at approximately +4 and +2, respectively, over the entire temperature range measured. Electron and X-ray diffraction studies reveal that the body-centered cubic symmetry (space group Im3̅, No. 204) is retained at temperatures as low as 100 K, indicating the absence of any types of charge-ordering in the charge-disproportionated CeCu3Fe4O12 phase. The magnetic susceptibility and neutron powder diffraction data illustrate that the antiferromagnetic ordering of Fe ions is predominant in the charge-disproportionated CeCu3Fe4O12 phase. These findings suggest that CeCu3Fe4O12 undergoes a new type of electronic phase in the ACu3Fe4O12 series and that the melting of the charge-ordering in CeCu3Fe4O12 is caused by the substantial decrease in the Fe valence and the resulting large deviation from the ideal abundance ratio of Fe(3+):Fe(5+) = 1:1 for rock-salt-type charge-ordering.


Assuntos
Cério/química , Cobre/química , Congelamento , Compostos de Ferro/química , Óxidos/química , Compostos de Ferro/síntese química , Modelos Moleculares , Estrutura Molecular , Óxidos/síntese química
14.
Inorg Chem ; 53(14): 7089-91, 2014 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-25000156

RESUMO

A novel perovskite oxide, CaCu3Rh4O12, has been synthesized under high-pressure and high-temperature conditions (15 GPa and 1273 K). Rietveld refinement of synchrotron X-ray powder diffraction data indicates that this compound crystallizes in a cubic AA'3B4O12-type perovskite structure. Synchrotron X-ray absorption and photoemission spectroscopy measurements reveal that the Cu and Rh valences are nearly trivalent. The spectroscopic analysis based on calculations suggests that the appropriate ionic model of this compound is Ca(2+)Cu(∼2.8+)3Rh(∼3.4+)4O12, as opposed to the conventional Ca(2+)Cu(2+)3Rh(4+)4O12. The uncommon valence state of this compound is attributed to the relative energy levels of the Cu 3d and Rh 4d orbitals, in which the large crystal-field splitting energy of the Rh 4d orbitals is substantial.


Assuntos
Compostos de Cálcio/síntese química , Óxidos/química , Compostos de Cálcio/química , Temperatura Alta , Óxidos/síntese química , Difração de Pó , Pressão , Titânio/química , Espectroscopia por Absorção de Raios X
15.
J Vet Med Sci ; 76(8): 1137-9, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24813669

RESUMO

We measured the plasma ghrelin and cortisol concentrations in non-lactating cows under fixed-time feeding conditions followed by an acute or gradual fasting treatment. During the 4 days before fasting, animals in Group 1 were fed a fixed amount of rations at 0800 and 1600 hr, and those in Group 2 were fed a gradually reduced amount. Thereafter, the plasma ghrelin concentrations of each animal were measured for 40 hr. The plasma ghrelin concentrations, which were low at the onset of fasting, increased before and after 0800 during fasting in Group 1, but not in Group 2. There were no significant differences in the plasma cortisol concentration within or between the groups. It was demonstrated that acute fasting induces elevation of the plasma ghrelin concentration, but that gradual fasting does not. This result suggests that fixed-time and fixed-quantity feeding caused a daily ghrelin rhythm in the cow and that this rhythm influenced changes in plasma ghrelin.


Assuntos
Jejum/fisiologia , Métodos de Alimentação/veterinária , Grelina/sangue , Hidrocortisona/sangue , Animais , Bovinos , Feminino , Fatores de Tempo
16.
BMC Plant Biol ; 14: 58, 2014 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-24602314

RESUMO

BACKGROUND: Flower color of soybean is primarily controlled by six genes, viz., W1, W2, W3, W4, Wm and Wp. This study was conducted to investigate the genetic and chemical basis of newly-identified flower color variants including two soybean mutant lines, 222-A-3 (near white flower) and E30-D-1 (light purple flower), a near-isogenic line (Clark-w4), flower color variants (T321 and T369) descended from the w4-mutable line and kw4 (near white flower, Glycine soja). RESULTS: Complementation tests revealed that the flower color of 222-A-3 and kw4 was controlled by the recessive allele (w4) of the W4 locus encoding dihydroflavonol 4-reductase 2 (DFR2). In 222-A-3, a single base was deleted in the first exon resulting in a truncated polypeptide consisting of 24 amino acids. In Clark-w4, base substitution of the first nucleotide of the fourth intron abolished the 5' splice site, resulting in the retention of the intron. The DFR2 gene of kw4 was not expressed. The above results suggest that complete loss-of-function of DFR2 gene leads to near white flowers. Light purple flower of E30-D-1 was controlled by a new allele at the W4 locus, w4-lp. The gene symbol was approved by the Soybean Genetics Committee. In E30-D-1, a single-base substitution changed an amino acid at position 39 from arginine to histidine. Pale flowers of T369 had higher expression levels of the DFR2 gene. These flower petals contained unique dihydroflavonols that have not yet been reported to occur in soybean and G. soja. CONCLUSIONS: Complete loss-of-function of DFR2 gene leads to near white flowers. A new allele of the W4 locus, w4-lp regulates light purple flowers. Single amino acid substitution was associated with light purple flowers. Flower petals of T369 had higher levels of DFR2 gene expression and contained unique dihydroflavonols that are absent in soybean and G. soja. Thus, mutants of the DFR2 gene have unique flavonoid compositions and display a wide variety of flower color patterns in soybean, from near white, light purple, dilute purple to pale.


Assuntos
Oxirredutases do Álcool/metabolismo , Flores/enzimologia , Flores/genética , Glycine max/enzimologia , Glycine max/genética , Proteínas de Plantas/metabolismo , Oxirredutases do Álcool/genética , Alelos , Flores/metabolismo , Pigmentação/genética , Pigmentação/fisiologia , Proteínas de Plantas/genética , Glycine max/metabolismo
17.
Biophys J ; 106(2): 421-9, 2014 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-24461017

RESUMO

Computer simulations have been demonstrated to be important for unraveling atomic mechanisms in biological systems. In this study, we show how combining unbiased molecular dynamic simulations with appropriate analysis tools can successfully describe metal-based drug interactions with DNA. To elucidate the noncovalent affinity of cisplatin's family to DNA, we performed extensive all-atom molecular dynamics simulations (3.7 µs total simulation length). The results show that the parent drug, cisplatin, has less affinity to form noncovalent adducts in the major groove than its aquo complexes. Furthermore, the relative position in which the drugs enter the major groove is dependent on the compound's net charge. Based on the simulations, we estimated noncovalent binding free energies through the use of Markov state models. In addition, and to overcome the lack of experimental information, we employed two additional methods: Molecular Mechanics Poisson-Boltzmann Surface Area (MMPB-SA) and steered molecular dynamics with the Jarzynski estimator, with an overall good agreement between the three methods. All complexes show interaction energies below 3 kcal/mol with DNA but the charged hydrolysis products have slightly more favorable binding free energies than the parent drug. Moreover, this study sets the precedent for future unbiased DNA-ligand simulations of more complex binders.


Assuntos
Antineoplásicos/metabolismo , Cisplatino/metabolismo , DNA/metabolismo , Simulação de Dinâmica Molecular , Antineoplásicos/química , Sequência de Bases , Cisplatino/química , DNA/química , DNA/genética , Cinética , Conformação de Ácido Nucleico , Termodinâmica
18.
J Chem Theory Comput ; 10(1): 282-8, 2014 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-26579911

RESUMO

Obtaining absolute binding free energies from unbiased ligand diffusion has attracted a significant amount of attention due to its implications in drug design. Several studies have used special purpose computers and software to achieve microsecond molecular dynamics which, combined with a Markov state model analysis, are capable of providing absolute binding free energies. We have recently developed a Monte Carlo based technique, PELE, capable of performing a dynamical exploration of the protein-ligand energy landscape including free ligand diffusion into the active site, at a fraction of the computational cost of molecular dynamics techniques. We demonstrate here the capabilities of our Monte Carlo technique in obtaining absolute binding free energies for a series of benzamidine like inhibitors into trypsin. Our results are in good agreement with experimental data and other molecular dynamics simulations, indicating that PELE can be a useful tool for quick estimates of binding free energies and mechanisms.

19.
Plant Mol Biol ; 84(3): 287-300, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24072327

RESUMO

There are substantial genotypic differences in the levels of flavonol glycosides (FGs) in soybean leaves. The first objective of this study was to identify and locate genes responsible for FG biosynthesis in the soybean genome. The second objective was to clone and verify the function of these candidate genes. Recombinant inbred lines (RILs) were developed by crossing the Kitakomachi and Koganejiro cultivars. The FGs were separated by high performance liquid chromatography (HPLC) and identified. The FGs of Koganejiro had rhamnose at the 6″-position of the glucose or galactose bound to the 3-position of kaempferol, whereas FGs of Kitakomachi were devoid of rhamnose. Among the 94 RILs, 53 RILs had HPLC peaks classified as Koganejiro type, and 41 RILs had peaks classified as Kitakomachi type. The segregation fitted a 1:1 ratio, suggesting that a single gene controls FG composition. SSR analysis, linkage mapping and genome database survey revealed a candidate gene in the molecular linkage group O (chromosome 10). The coding region of the gene from Koganejiro, designated as GmF3G6″Rt-a, is 1,392 bp long and encodes 464 amino acids, whereas the gene of Kitakomachi, GmF3G6″Rt-b, has a two-base deletion resulting in a truncated polypeptide consisting of 314 amino acids. The recombinant GmF3G6″Rt-a protein converted kaempferol 3-O-glucoside to kaempferol 3-O-rutinoside and utilized 3-O-glucosylated/galactosylated flavonols and UDP-rhamnose as substrates. GmF3G6″Rt-b protein had no activity. These results indicate that GmF3G6″Rt encodes a flavonol 3-O-glucoside (1 â†’ 6) rhamnosyltransferase and it probably corresponds to the Fg2 gene. GmF3G6″Rt was designated as UGT79A6 by the UGT Nomenclature Committee.


Assuntos
Mapeamento Cromossômico , Genes de Plantas , Glycine max/genética , Hexosiltransferases/genética , Proteínas de Soja/genética , Sequência de Aminoácidos , Sequência de Bases , Cromatografia Líquida de Alta Pressão , Clonagem Molecular , Primers do DNA , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Hexosiltransferases/química , Hexosiltransferases/isolamento & purificação , Dados de Sequência Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Homologia de Sequência de Aminoácidos , Proteínas de Soja/química , Proteínas de Soja/isolamento & purificação
20.
Inorg Chem ; 52(23): 13751-61, 2013 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-24224928

RESUMO

Unusual electronic phase transitions in the A-site ordered perovskites LnCu3Fe4O12 (Ln: trivalent lanthanide ion) are investigated. All LnCu3Fe4O12 compounds are in identical valence states of Ln(3+)Cu(2+)3Fe(3.75+)4O12 at high temperature. LnCu3Fe4O12 with larger Ln ions (Ln = La, Pr, Nd, Sm, Eu, Gd, Tb) show an intersite charge transfer transition (3Cu(2+) + 4Fe(3.75+) → 3Cu(3+) + 4Fe(3+)) in which the transition temperature decreases from 360 to 240 K with decreasing Ln ion size. In contrast, LnCu3Fe4O12 with smaller Ln ions (Ln = Dy, Ho, Er, Tm Yb, Lu) transform into a charge-disproportionated (8Fe(3.75+) → 5Fe(3+) + 3Fe(5+)) and charge-ordered phase below ∼250-260 K. The former series exhibits metal-to-insulator, antiferromagnetic, and isostructural volume expansion transitions simultaneously with intersite charge transfer. The latter shows metal-to-semiconductor, ferrimagnetic, and structural phase transitions simultaneously with charge disproportionation. Bond valence calculation reveals that the metal-oxygen bond strains in these compounds are classified into two types: overbonding or compression stress (underbonding or tensile stress) in the Ln-O (Fe-O) bond is dominant in the former series, while the opposite stresses or bond strains are found in the latter. Intersite charge transfer transition temperatures are strongly dependent upon the global instability indices that represent the structural instability calculated from the bond valence sum, whereas the charge disproportionation occurs at almost identical temperatures, regardless of the magnitude of structural instability. These findings provide a new aspect of the structure-property relationship in transition metal oxides and enable precise control of electronic states by bond strains.


Assuntos
Compostos de Cálcio/química , Compostos de Ferro/química , Elementos da Série dos Lantanídeos/química , Óxidos/química , Titânio/química , Temperatura Baixa , Cristalografia por Raios X , Elétrons , Modelos Moleculares , Transição de Fase
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