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1.
Nanotechnology ; 27(49): 494001, 2016 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-27823991

RESUMO

Shifts from the expected nuclear magnetic resonance frequencies of antimony and bismuth donors in silicon of greater than a megahertz are observed in electrically detected magnetic resonance spectra. Defects created by ion implantation of the donors are discussed as the source of effective electric field gradients generating these shifts via quadrupole interaction with the nuclear spins. The experimental results are modeled quantitatively by molecular orbital theory for a coupled pair consisting of a donor and a spin-dependent recombination readout center.

2.
Nano Lett ; 15(6): 3885-93, 2015 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-25993500

RESUMO

The introduction of stable isotopes in the fabrication of semiconductor nanowires provides an additional degree of freedom to manipulate their basic properties, design an entirely new class of devices, and highlight subtle but important nanoscale and quantum phenomena. With this perspective, we report on phonon engineering in metal-catalyzed silicon nanowires with tailor-made isotopic compositions grown using isotopically enriched silane precursors (28)SiH4, (29)SiH4, and (30)SiH4 with purity better than 99.9%. More specifically, isotopically mixed nanowires (28)Si(x)(30)Si(1-x) with a composition close to the highest mass disorder (x ∼ 0.5) were investigated. The effect of mass disorder on the phonon behavior was elucidated and compared to that in isotopically pure (29)Si nanowires having a similar reduced mass. We found that the disorder-induced enhancement in phonon scattering in isotopically mixed nanowires is unexpectedly much more significant than in bulk crystals of close isotopic compositions. This effect is explained by a nonuniform distribution of (28)Si and (30)Si isotopes in the grown isotopically mixed nanowires with local compositions ranging from x = ∼0.25 to 0.70. Moreover, we also observed that upon heating, phonons in (28)Si(x)(30)Si(1-x) nanowires behave remarkably differently from those in (29)Si nanowires suggesting a reduced thermal conductivity induced by mass disorder. Using Raman nanothermometry, we found that the thermal conductivity of isotopically mixed (28)Si(x)(30)Si(1-x) nanowires is ∼30% lower than that of isotopically pure (29)Si nanowires in agreement with theoretical predictions.


Assuntos
Nanofios/química , Fônons , Silício/química , Silanos/química
3.
ScientificWorldJournal ; 2014: 457350, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24715812

RESUMO

Three species of otter can be found throughout Malay Peninsula: Aonyx cinereus, Lutra sumatrana, and Lutrogale perspicillata. In this study, we focused on the A. cinereus population that ranges from the southern and the east coast to the northern regions of Malay Peninsula up to southern Thailand to review the relationships between the populations based on the mitochondrial D-loop region. Forty-eight samples from six populations were recognized as Johor, Perak, Terengganu, Kelantan, Ranong, and Thale Noi. Among the 48 samples, 33 were identified as A. cinereus, seven as L. sumatrana, and eight as L. perspicillata. Phylogenetically, two subclades formed for A. cinereus. The first subclade grouped all Malay Peninsula samples except for samples from Kelantan, and the second subclade grouped Kelantan samples with Thai sample. Genetic distance analysis supported the close relationships between Thai and Kelantan samples compared to the samples from Terengganu and the other Malaysian states. A minimum-spanning network showed that Kelantan and Thailand formed a haplogroup distinct from the other populations. Our results show that Thai subspecies A. cinereus may have migrated to Kelantan from Thai mainland. We also suggest the classification of a new subspecies from Malay Peninsula, the small-clawed otter named A. cinereus kecilensis.


Assuntos
DNA/genética , Fezes , Lontras/classificação , Animais , Malásia , Lontras/genética , Tailândia
4.
Phys Rev Lett ; 104(13): 137402, 2010 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-20481913

RESUMO

As the deepest group-V donor in Si, Bi has by far the largest hyperfine interaction and also a large I = 9/2 nuclear spin. At zero field this splits the donor ground state into states having total spin 5 and 4, which are fully resolved in the photoluminescence spectrum of Bi donor bound excitons. Under a magnetic field, the 60 expected allowed transitions cannot be individually resolved, but the effects of the nuclear spin distribution, -9/2 < or = I(z) < or = 9/2, are clearly observed. A strong hyperpolarization of the nuclear spin towards I(z) = -9/2 is observed to result from the nonresonant optical excitation. This is very similar to the recently reported optical hyperpolarization of P donors observed by EPR at higher magnetic fields. We introduce a new model to explain this effect, and predict that it may be very fast.

5.
Science ; 364(6437): 252, 2019 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-30890587

RESUMO

The near-Earth carbonaceous asteroid 162173 Ryugu is thought to have been produced from a parent body that contained water ice and organic molecules. The Hayabusa2 spacecraft has obtained global multicolor images of Ryugu. Geomorphological features present include a circum-equatorial ridge, east-west dichotomy, high boulder abundances across the entire surface, and impact craters. Age estimates from the craters indicate a resurfacing age of [Formula: see text] years for the top 1-meter layer. Ryugu is among the darkest known bodies in the Solar System. The high abundance and spectral properties of boulders are consistent with moderately dehydrated materials, analogous to thermally metamorphosed meteorites found on Earth. The general uniformity in color across Ryugu's surface supports partial dehydration due to internal heating of the asteroid's parent body.

6.
Scanning ; 30(4): 347-53, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18615457

RESUMO

We present advanced semiconductor diagnosis by using electron-beam-induced current (EBIC) technique. By varying the parameters such as temperature, accelerating voltage (V(acc)), bias voltage, and stressing time, it is possible to extend EBIC application from conventional defect characterization to advanced device diagnosis. As an electron beam can excite a certain volume even beneath the surface passive layer, EBIC can be effectively employed to diagnose complicated devices with hybrid structure. Three topics were selected to demonstrate EBIC applications. First, the recombination activities of grain boundaries and their interaction with Fe impurity in photovoltaic multicrystalline Si (mc-Si) are clarified by temperature-dependent EBIC. Second, the detection of dislocations between strained-Si and SiGe virtual substrate are shown to overcome the limitation of depletion region. Third, the observation of leakage sites in high-k gate dielectric is demonstrated for the characterization of advanced hybrid device structures.

7.
Acta Biol Hung ; 59 Suppl: 237-9, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18652397

RESUMO

Ci-TK and Ci-TK-R are authentic tachykinin (TK) and TK receptor isolated from a protochordate, Ciona intestinalis. In this study, we investigated a novel function of TK as an enhancer of oocyte growth. Ci-TK-R is expressed specifically in the Ciona vitellogenic oocytes. Moreover, administration of Ci-TK to the Ciona ovary resulted in upregulation of gene expression and enzymatic activity of several proteases. Moreover, maturation of the Ciona oocytes from the vitellogenic stage to the post-vitellogenic stage was induced in the presence of Ci-TK, which was completely blocked by addition of protease inhibitors.


Assuntos
Ciona intestinalis/metabolismo , Neuropeptídeos/metabolismo , Receptores de Neuropeptídeos/metabolismo , Animais , Ciona intestinalis/genética , Evolução Molecular , Feminino , Neuropeptídeos/genética , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Receptores de Taquicininas/genética , Receptores de Taquicininas/metabolismo , Taquicininas/genética , Taquicininas/metabolismo , Vertebrados
8.
J Clin Invest ; 65(2): 256-67, 1980 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-7356677

RESUMO

We investigated the mechanism of gastroesophageal reflux (GER) in 10 health volunteer subjects. Continuous recordings of intraluminal esophageal pH and pressure were obtained on two consecutive nights from 6:00 p.m. to 6:30 a.m. in each subject. During each study, the subject remained recumbent, except to eat a standardized meal after 1 h of basal recording. A manometric assembly with seven recording lumens monitored: (a) lower esophageal sphincter (LES) pressure via a sleeve device 6.5 cm in length, (b) esophageal-body motor activity, (c) swallowing activity in the pharynx, and (d) gastric pressure. An electrode 5 cm above the LES recorded esophageal pH. Sleep was monitored by electroencephalogram. All subjects showed wide variations of basal LES pressure. GER was not related to low steady-state basal LES pressure, but rather occurred during transient 5-30 s episodes of inappropriate complete LES relaxation. The inappropriate LES relaxations were usually either spontaneous or immediately followed appropriate sphincter relaxation induced by swallowing. The majority of GER episodes occurred within the first 3 h after eating. During the night LES relaxation and GER occurred only during transient arousals from sleep or when the subjects were fully awake, but not during stable sleep. After GER the esophagus was generally cleared of refluxed acid by primary peristalsis and less frequently by secondary peristalsis. Nonperistaltic contractions were less effective than peristalsis for clearing acid from the esophagus. We conclude that in asymptomatic recumbent subjects GER is related to transient inappropriate LES relaxations rather than to low steady-state basal LES pressure and also, that primary perstalsis is the major mechanism that clears the esophagus of refluxed material.


Assuntos
Junção Esofagogástrica/fisiopatologia , Refluxo Gastroesofágico/etiologia , Adulto , Deglutição , Refluxo Gastroesofágico/fisiopatologia , Humanos , Concentração de Íons de Hidrogênio , Peristaltismo , Postura , Pressão , Fases do Sono
9.
Mol Cell Biol ; 11(6): 3317-25, 1991 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-2038334

RESUMO

The human CCG1 gene complements tsBN462, a temperature-sensitive G1 mutant of the BHK21 cell line. The previously cloned cDNA turned out to be a truncated form of the actual CCG1 cDNA. The newly cloned CCG1 cDNA was 6.0 kb and encoded a protein with a molecular mass of 210 kDa. Using an antibody to a predicted peptide from the CCG1 protein, a protein with a molecular mass of over 200 kDa was identified in human, monkey, and hamster cell lines. In the newly defined C-terminal region, an acidic domain was found. It contained four consensus target sequences for casein kinase II and was phosphorylated by this enzyme in vitro. However, this C-terminal region was not required to complement tsBN462 mutation since the region encoding the C-terminal part was frequently missing in complemented clones derived by DNA-mediated gene transfer. CCG1 contains a sequence similar to the putative DNA-binding domain of HMG1 in addition to the previously detected amino acid sequences common in nuclear proteins, such as a proline cluster and a nuclear translocation signal. Consistent with these predictions, CCG1 was present in nuclei, possessed DNA-binding activity, and was eluted with similar concentrations of salt, 0.3 to 0.4 M NaCl either from isolated nuclei or from a DNA-cellulose column.


Assuntos
Proteínas de Ligação a DNA/genética , Fase G1 , Proteínas Nucleares/genética , Fatores Associados à Proteína de Ligação a TATA , Fator de Transcrição TFIID , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , Clonagem Molecular , Códon/genética , DNA/genética , DNA/isolamento & purificação , Proteínas de Ligação a DNA/isolamento & purificação , Proteínas de Ligação a DNA/metabolismo , Escherichia coli/genética , Histona Acetiltransferases , Humanos , Dados de Sequência Molecular , Peso Molecular , Proteínas Nucleares/isolamento & purificação , Proteínas Nucleares/metabolismo , Sondas de Oligonucleotídeos , Reação em Cadeia da Polimerase , Proteínas Recombinantes/isolamento & purificação , Homologia de Sequência do Ácido Nucleico , Temperatura
10.
Mol Cell Biol ; 10(2): 577-84, 1990 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-2300055

RESUMO

At the nonpermissive temperature, premature chromosome condensation (PCC) occurs in tsBN2 cells derived from the BHK cell line, which can be converted to the Ts+ phenotype by the human RCC1 gene. To prove that the RCC1 gene is the mutant gene in tsBN2 cells, which have RCC1 mRNA and protein of the same sizes as those of BHK cells, RCC1 cDNAs were isolated from BHK and tsBN2 cells and sequenced to search for mutations. The hamster (BHK) RCC1 cDNA encodes a protein of 421 amino acids homologous to the human RCC1 protein. In a comparison of the base sequences of BHK and BN2 RCC1 cDNAs, a single base change, cytosine to thymine (serine to phenylalanine), was found in the 256th codon of BN2 RCC1 cDNA. The same transition was verified in the RCC1 genomic DNA by the polymerase chain reaction method. BHK RCC1 cDNA, but not tsBN2 RCC1 cDNA, complemented the tsBN2 mutation, although both have the same amino acid sequence except for one amino acid at the 256th codon. This amino acid change, serine to phenylalanine, was estimated to cause a profound structural change in the RCC1 protein.


Assuntos
Proteínas de Ciclo Celular , Ciclo Celular , Cromossomos/fisiologia , Proteínas de Ligação a DNA/genética , Genes , Fatores de Troca do Nucleotídeo Guanina , Mutação , Proteínas Nucleares , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , Clonagem Molecular , Cricetinae , DNA/genética , Biblioteca Gênica , Humanos , Rim , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Sondas de Oligonucleotídeos , Fenótipo , Reação em Cadeia da Polimerase , Conformação Proteica , Mapeamento por Restrição , Homologia de Sequência do Ácido Nucleico , Temperatura
11.
Mol Cell Biol ; 13(10): 6367-74, 1993 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-8413235

RESUMO

The tsBN7 cell line, one of the mutant lines temperature sensitive for growth which have been isolated from the BHK21 cell line, was found to die by apoptosis following a shift to the nonpermissive temperature. The induced apoptosis was inhibited by a protein synthesis inhibitor, cycloheximide, but not by the bcl-2-encoded protein. By DNA-mediated gene transfer, we cloned a cDNA that complements the tsBN7 mutation. It encodes a novel hydrophobic protein, designated DAD1, which is well conserved (100% identical amino acids between humans and hamsters). By comparing the base sequences of the parental BHK21 and tsBN7 DAD1 cDNAs, we found that the DAD1-encoding gene is mutated in tsBN7 cells. The DAD1 protein disappeared in tsBN7 cells following a shift to the nonpermissive temperature, suggesting that loss of the DAD1 protein triggers apoptosis.


Assuntos
Apoptose/fisiologia , Proteínas de Membrana , Proteínas/genética , Proteínas/fisiologia , Proteínas de Xenopus , Sequência de Aminoácidos , Animais , Apoptose/genética , Proteínas Reguladoras de Apoptose , Sequência de Bases , Linhagem Celular , Linhagem Celular Transformada , Clonagem Molecular , Sequência Conservada , Cricetinae , Cicloeximida/farmacologia , DNA , Teste de Complementação Genética , Células HeLa , Humanos , Dados de Sequência Molecular , Homologia de Sequência de Aminoácidos , Temperatura , Xenopus laevis
12.
J Nanosci Nanotechnol ; 7(9): 3323-7, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18019168

RESUMO

Low-energy cathodoluminescence (CL) imaging and spectroscopy technique was employed to study the impurity distribution in individual ZnO hexagonal nanotubes fabricated by metalorganic chemical vapor deposition on the sapphire (0001) substrate. The CL spectra at 10 K show that acceptor and donor impurities are incorporated in the ZnO nanotubes. CL monochromatic images indicate that the concentration of donor is higher at the bottom part and the distribution of acceptors is more inhomogeneous at the surface of the nanotubes. The non-uniform defects and impurities distributions are explained by unstable growth conditions and contamination from the environment. These results indicate that the low-energy CL is a very powerful method to investigate the inhomogeneity of luminescence properties in the individual nanostructures.


Assuntos
Nanopartículas Metálicas/química , Nanotecnologia/métodos , Óxido de Zinco/química , Óxido de Alumínio , Química Orgânica/métodos , Eletrodos , Elétrons , Luminescência , Microscopia Eletrônica de Varredura , Nanopartículas/química , Nanoestruturas/química , Nanotubos , Nanotubos de Carbono/química , Fótons , Temperatura
13.
Lab Chip ; 17(8): 1481-1486, 2017 04 11.
Artigo em Inglês | MEDLINE | ID: mdl-28301613

RESUMO

This paper presents the formation of complex cross-sectional microfibers using three-dimensional microfluidic devices. The compartments and shapes of core and shell layers in the microfibers were independently controlled via three-dimensional fluidic channels fabricated by the combination of sheath units. The number of layers is easily expanded by the stacking of these units. Therefore, the highly heterogeneous microfibers of alginate hydrogel are obtained in polydimethylsiloxane structures. This widely expandable method has great potential for the development of functional and complex fiber-shaped materials.

14.
Oncogene ; 16(3): 369-80, 1998 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-9467962

RESUMO

p21cip1/waf1/sdi1 is a universal cyclin-Cdk kinase inhibitor that has two functional domains; one binds and inhibits cyclin-Cdk activity and the other binds PCNA and thereby inhibits elongation by DNA polymerase. When transiently expressed in hamster BHK21 cells we found that human p21 was able to cause cell cycle arrest in G1 phase; this arrest was counteracted by coexpression of E2F-1 or SV40 large T antigen. To study the effect of p21 overexpression in vivo, BHK21 cell clones inducibly expressing human p21 (Tet-p21) driven by the tetracycline (Tet)-repressible promoter were established. The maximum induced p21 levels in the absence of Tet were estimated to be ten times that of endogenous hamster p21. As p21 levels rose following removal of Tet, p21-associated histone H1 kinase activity was increased and concomitantly cell growth and DNA synthesis were reduced. Tet-p21 BHK21 cells became arrested in G1 phase and lost colony forming ability irreversibly 2-4 days after removal of Tet. The induction of cyclin E- and cyclin A-associated kinase activities was diminished when G0-synchronized Tet-p21 BHK21 cells were serum stimulated in the absence of Tet. Increased binding of p21 to PCNA and cyclin D1-Cdk4 was detected in induced cells. Overexpression of p21 led to cell death in BHK21 cells at 39.5 degrees C within 4 days.


Assuntos
Quinases relacionadas a CDC2 e CDC28 , Quinases Ciclina-Dependentes/antagonistas & inibidores , Ciclinas/fisiologia , Inibidores Enzimáticos/farmacologia , Proteínas Proto-Oncogênicas , Animais , Antibacterianos/farmacologia , Proteína Quinase CDC2/metabolismo , Morte Celular , Divisão Celular , Linhagem Celular , Sobrevivência Celular , Cricetinae , Ciclina D1/metabolismo , Quinase 2 Dependente de Ciclina , Quinase 4 Dependente de Ciclina , Inibidor de Quinase Dependente de Ciclina p21 , Quinases Ciclina-Dependentes/metabolismo , Ciclinas/biossíntese , Ciclinas/genética , Expressão Gênica/efeitos dos fármacos , Humanos , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Quinases/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Tetraciclina/farmacologia
15.
Oncogene ; 18(10): 1797-806, 1999 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-10086334

RESUMO

tsBN462 cells, which have a point mutation in CCG1/TAF(II)250, a component of TFIID complex, arrest in G1 at the nonpermissive temperature of 39.5 degrees C. Overexpression of D-type cyclins rescued the cell cycle arrest of tsBN462 cells, suggesting that the cell cycle arrest was through Rb. Consistent with this, overexpression of E2F-1, whose function is repressed by the hypophosphorylated form of Rb, also rescued the cell cycle arrest. Moreover, expression of the viral oncoproteins SV40 large T antigen and HPV16 E7, which both bind Rb and inactivate its function, rescued the cell cycle arrest, whereas HPV16 E6 did not. Mutation of the Rb-binding motif in E7 abrogated its ability to rescue the cell cycle arrest. Expression of exogenous cyclin D1, SV40 large T antigen or CCG1/TAF(II)250 increased cyclin A expression at 39.5 degrees C. Coexpression of HPV16 E7 and adenovirus E1b19K, which blocks apoptosis, rescued the proliferation of tsBN462 cells at 38.5 degrees C. To investigate the mechanism underlying the lack of cyclin D1 expression, deletion analysis of cyclin D1 promoter was performed. The 0.15 kbp cyclin D1 core promoter region, which lacks any transcription factor binding motifs, still exhibited a temperature-sensitive phenotype in tsBN462 cells suggesting that CCG1/TAF(II)250 is critical for the function of the cyclin D1 core promoter.


Assuntos
Antígenos Transformantes de Poliomavirus/metabolismo , Proteínas de Transporte , Proteínas de Ciclo Celular , Ciclina D1/metabolismo , Proteínas de Ligação a DNA/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Oncogênicas Virais/metabolismo , Fatores Associados à Proteína de Ligação a TATA , Fator de Transcrição TFIID , Fatores de Transcrição/metabolismo , Animais , Antígenos Transformantes de Poliomavirus/genética , Ciclo Celular , Divisão Celular , Linhagem Celular , Cricetinae , Ciclina D1/genética , Proteínas de Ligação a DNA/genética , Fatores de Transcrição E2F , Fator de Transcrição E2F1 , Fase G1 , Histona Acetiltransferases , Humanos , Mutagênese , Proteínas Nucleares/genética , Proteínas Oncogênicas Virais/genética , Proteínas E7 de Papillomavirus , Fenótipo , Regiões Promotoras Genéticas , Proteína 1 de Ligação ao Retinoblastoma , Temperatura , Fator de Transcrição DP1
16.
Biochim Biophys Acta ; 788(3): 267-73, 1984 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-6087910

RESUMO

The beta-isopropylmalate dehydrogenase (2-hydroxy-4-methyl-3-carboxyvalerate: NAD+ oxidoreductase, EC 1.1.1.85) gene from Baccilus coagulans was cloned and expressed in Escherichia coli C600, using pBR322 as a vector plasmid. The B. coagulans enzyme was purified to a homogeneous state from the E. coli carrying a pBR322 - the B. coaglulans enzyme gene hybrid plasmid. The enzyme consists of two subunits of equal molecular weight (4.4 X 10(4) ). The enzyme activity was stimulated by 0.5 mM Mn2+, Mg2+ and Co2+. The enzyme was strongly inhibited by 0.2 mM p-chloromercuribenzoate and the inhibition was completely recovered by 1 mM dithiothreitol. The B. coagulans enzyme was thermostabilized by 1.5 M NaCl. The B. coagulans enzyme is a composite of alpha-helix, beta-sheet and remainder. The secondary structure of the enzyme was appreciably altered by 0.5 mM MgCl2 and 1.5 M NaCl.


Assuntos
Oxirredutases do Álcool/genética , Bacillus/enzimologia , Clonagem Molecular , Escherichia coli/enzimologia , 3-Isopropilmalato Desidrogenase , Oxirredutases do Álcool/isolamento & purificação , Cátions/farmacologia , Enzimas de Restrição do DNA/metabolismo , Temperatura Alta , Hibridização de Ácido Nucleico , Plasmídeos , Conformação Proteica
17.
Biochim Biophys Acta ; 708(3): 300-4, 1982 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-6816286

RESUMO

The thermostability of the isozymes from pig heart (H) and muscle (M) lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 1.1.1.27) decreased in the order of H4 greater than M4 greater than H3M greater than HM3 greater than H2M2, while the thermostability of the isozymes from guanidinated H4 and M4 increased as guanidinated H monomer was substituted by M monomer. The increased thermostability of H4 increased as guanidinated H monomer was substituted by M monomer. The increased thermostability of H4 on guanidination of five lysine residues per subunit was due to the decrease in the standard activation entropy, and no change in the standard activation enthalpy was observed. The more increased thermostability of H4 on further guanidination of lysine residues from 5 to 15 per subunit was due to the increase of the standard activation enthalpy which overcame the decrease in thermostability due to the increase of the standard activation entropy. The results indicate two different mechanisms of stabilization depending on the degree of guanidination. The increase of thermostability, as measured by the change of the standard activation free energy for thermal inactivation of H2M2, was almost the same as that of H4 on guanidination of five lysine residues in an H monomer. This result and the order of thermostability of the isozymes from unmodified and guanidinated H4 and M4 suggest that the increase of thermostability of hybrid isozymes on guanidination of H monomer is due to the change of the heterologous subunit interactions.


Assuntos
Guanidinas/farmacologia , L-Lactato Desidrogenase/metabolismo , Animais , Estabilidade de Medicamentos , Guanidina , Temperatura Alta , Isoenzimas , Cinética , Lisina , Substâncias Macromoleculares , Multimerização Proteica , Suínos , Termodinâmica
18.
Biochim Biophys Acta ; 1326(1): 138-48, 1997 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-9188809

RESUMO

Purple membrane (PM) shows denaturation when spread over an air/water interface. We established a technique, which we call the spectroscopic surface denaturation quantifying (SSDQ) technique, that uses infrared linear dichroism to determine the amount of native structural bacteriorhodopsin (BR) in PM Langmuir-Blodgett (LB) films. Using the SSDQ technique we found that the conformational change after surface denaturation of BR was the same as that caused by ethanol treatment. By extrapolating the data of the amount of non-denatured BR molecules in PM LB films vs. the area of a single BR molecule on an air/water interface, we also found that the surface area of a single non-denatured BR molecule was 11.5 nm2, which is consistent with that determined by high-resolution electron cryo-microscopy and electron diffraction (EMD). These results demonstrate that the SSDQ technique is effective in quantifying the amount of native structural BR in PM LB films. The SSDQ technique is also applicable to other types of protein consisting of alpha-helical conformation.


Assuntos
Bacteriorodopsinas/análise , Membrana Purpúrea/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Bacteriorodopsinas/ultraestrutura , Matemática , Conformação Proteica
19.
Biochim Biophys Acta ; 581(2): 334-41, 1979 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-518917

RESUMO

Acetamidination of pig heart lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 1.1.1.27) with ethyl acetimidate resulted in an increase of thermostability, and covalent bridge formation between pairs of lysine residues is observed. Guanidination with O-methylisourea of the enzyme also increases the thermostability, but such a bridge seems not to be formed. Increased thermostability of guanidinated enzyme is considered to be due to the shift of the pK values of the lysine residues from 10.5 to 12.5 after guanidination. Modification experiments with carbodiimide reveals that the enzyme contains 4.6 pairs of neighboring lysine and carboxyl residues per subunit, and amide bonding between 3.2 pairs results in an increase of thermostability. Guanidination of 4.6 Lys/subunit of the enzyme yields an enzyme derivative with considerably increased thermostability. Salt bridge formation between the 4.6 pairs of neighboring carboxyl and guanidinated lysine residues per subunit might make a major contribution to the increased thermostability of the guanidinated enzyme.


Assuntos
L-Lactato Desidrogenase , Compostos de Metilureia , Miocárdio/enzimologia , Animais , Carbodi-Imidas , Estabilidade de Medicamentos , Guanidinas , Temperatura Alta , Imidoésteres , Cinética , L-Lactato Desidrogenase/metabolismo , Lisina , Ligação Proteica , Conformação Proteica , Suínos
20.
Biochim Biophys Acta ; 1474(3): 299-308, 2000 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-10779681

RESUMO

In an experiment in which rats were allowed free access to food and water, the rats did not eat the diet containing a mushroom Pleurotus ostreatus even if they were emaciated. A P. ostreatus lectin (POL) was isolated from the mushroom as the food intake-suppression principle. In hemagglutination inhibition assays, Me-alphaGalNAc was the most potent inhibitor among the monosaccharides tested. Among all the sugars tested, 2'-fucosyllactose (Fucalpha1-->2Galbeta1-->4Glc) was the strongest inhibitor and its inhibitory potency was five times greater than that of Me-alphaGalNAc. POL exhibited a binding ability to bovine submaxillary mucin (BSM) and asialo-BSM and the other glycoproteins were inert to the binding. The food intake-suppressing activity of POL was dependent on the dose. The diet containing 0.1% POL caused a 50% decrease in the food intake of rats against the control.


Assuntos
Depressores do Apetite/isolamento & purificação , Lectinas/isolamento & purificação , Pleurotus/química , Aminoácidos/análise , Animais , Depressores do Apetite/farmacologia , Cátions , Cromatografia por Troca Iônica , Dieta , Ingestão de Alimentos/efeitos dos fármacos , Eletroforese em Gel de Poliacrilamida , Testes de Hemaglutinação , Temperatura Alta , Concentração de Íons de Hidrogênio , Focalização Isoelétrica , Lectinas/química , Lectinas/farmacologia , Masculino , Metais/farmacologia , Extratos Vegetais/farmacologia , Ratos , Ratos Wistar , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
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