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101.
Summary Polypeptides, synthesized from a mixture of amino acid amides by microwave heating during repeated hydration-dehydration cycles, showed hydrolase- and oxidoreductase-like catalytic activities. Poly(GAVDH), polypeptides synthesized from an equimolar mixture (each 0.1 M) of glycinamide,l-alaninamide,l-valinamide,l-aspartic acid -amide, andl-histidinamide, catalyzed the hydrolysis of PNPAc. The hydrolytic rate of PNPAc with poly(GAVDH) was the quadruple of that ofl-histidine alone. Though the kcat values of different resulting polypeptides were 103–106 times less than those of native hydrolases, the Km value of the polypeptides further containing phenylalanine residues was nearly equal to that of the esterase. This result indicates the presence of hydrophobic interaction between a substrate and the polypeptides. Resulting polypeptides also showed catalytic activity for the reduction of ferricyanide ion [Fe(CN)3–] with NADH. The polypeptides seemed to have a strong affinity for adenine nucleotides, because the reaction was inhibited by adenine derivatives such as NAD+ and AppA. A hypothesis for the emergence of primitive protein enzymes is discussed. 相似文献
102.
Nodules of cowpea plants (Vigna unguiculata (L.) Walp. cv. Vita 3 :Bradyrhizobium CB756) cultured for periods of 23 d with their root systems maintained in atmospheres containing a range of partial pressures
of O2 (pO2; 1–80%, v/v, in N2) formed and exported ureides (allantoin and allantoic acid) as the major products of fixation at all pO2 tested. In sub-ambient pO2 (1 and 2.5%) nodules contained specific activities of uricase (urate: O2 oxidoreductase; EC 1.7.3.3) and allantoinase (allantoin hydrolyase; EC 3.5.2.5) as much as sevenfold higher than in those
from air. On a cell basis, uninfected cells in nodules from 1% O2 contained around five times the level of uricase. Except for NAD: glutamate synthase (EC 1.4.1.14), which was reduced in
sub-ambient O2, the activities of other enzymes of ureide synthesis were relatively unaffected by pO2. Short-term effects of pO2 on assimilation of fixed nitrogen were measured in nodules of air-grown plants exposed to subambient pO2 (1, 2.5 or 5%, v/v in N2) and15N2. Despite a fall in total15N2 fixation, ureide synthesis and export was maintained at a high level except in 1% O2 where formation was halved. The data indicate that in addition to the structural and diffusional adaptations of cowpea nodules
which allow the balance between O2 supply and demand to be maintained over a wide range of pO2, nodules also show evidence of biochemical adaptations which maintain and enhance normal pathways for the assimilation of
fixed nitrogen.
This work was supported by a grant from the Australian Research Council (to C.A.A.) and an Australian Development Assistance
Bureau postgraduate fellowship (to F.D.D.). 相似文献
103.
In the last few years our knowledge of the structure and function of Photosystem II in oxygen-evolving organisms has increased significantly. The biochemical isolation and characterization of essential protein components and the comparative analysis from purple photosynthetic bacteria (Deisenhofer, Epp, Miki, Huber and Michel (1984) J Mol Biol 180: 385–398) have led to a more concise picture of Photosystem II organization. Thus, it is now generally accepted that the so-called D1 and D2 intrinsic proteins bind the primary reactants and the reducing-side components. Simultaneously, the nature and reaction kinetics of the major electron transfer components have been further clarified. For example, the radicals giving rise to the different forms of EPR Signal II have recently been assigned to oxidized tyrosine residues on the D1 and D2 proteins, while the so-called Q400 component has been assigned to the ferric form of the acceptor-side iron. The primary charge-separation has been meaured to take place in about 3 ps. However, despite all recent major efforts, the location of the manganese ions and the water-oxidation mechanism still remain largely unknown. Other topics which lately have received much attention include the organization of Photosystem II in the thylakoid membrane and the role of lipids and ionic cofactors like bicarbonate, calcium and chloride. This article attempts to give an overall update in this rapidly expanding field. 相似文献
104.
F_(IO_2)(吸入气氧浓度)为12.35、9.87及7.7l%,分别吸入10、8及5min时,心功能呈代偿性增强改变。F_(IO_2)为9.37%、吸入20min时心功能的变化趋势与9.87%8min时仍基本相同。继发性缺二氧化碳对缺氧引起的心功能代偿性增强,在一定程度上起抵消作用。F_(IO_2)为9.87%时的缺氧程度约相当于18km高空加压供氧总压值为15.3kPa(115mmHg)时的缺氧。单纯从缺氧因素考虑,将总压值由常用的17.3kPa(130mmHg)降低为15.3kPa是可允许的。 相似文献
105.
Cora-Jean S. Edgell Jill E. Haizlip C. Robert Bagnell Joan P. Packenham Paul Harrison Barry Wilbourn Victoria J. Madden 《In vitro cellular & developmental biology. Plant》1990,26(12):1167-1172
Summary Weibel-Palade bodies are ultrastructurally defined organelles found only in vascular endothelial cells. Because endothelium
in corpo is very dispersed, isolation and further characterization of this organelle has been dependent on increasing the
number of cells in culture. However, primary isolates of endothelial cells have a limited replication potential and tend to
senesce in culture. In this report, EA.hy926, a continuously replicating cell line derived from human endothelium, is shown
to contain Weibel-Palade bodies. Electron micrographs demonstrate the ultrastructural characteristics of these tissue-specific
organelles and their cytoplasmic distribution in EA.hy926 cells. Von Willebrand factor, which has been shown to exist in Weibel
Palade bodies, is demonstrated by immunofluorescence in discrete rod-shaped organelles whose size, shape, and distribution
are consistent with that of Weibel-Palade bodies in primary endothelial cell cultures. Rapid release of von Willebrand factor
can be induced by calcium ionophore, and large multimeric forms of the protein are found in EA.hy926 cells. These two properties
are consistent with the function currently ascribed to Weibel Palade bodies: storage of multimerized von Willebrand factor.
Thus ultrastructural, immunologic, and functional data establish the existence of this as yet poorly understood tissue-specific
organelle in a continuous, vigorously replicating human cell line. 相似文献
106.
Patricia A. McLenachan Peter J. Lockhart H. Rick Faber Brian C. Mansfield 《Journal of molecular evolution》1996,42(2):273-280
The pregnancy-specific 1-glycoproteins (PSG) form a large family of closely related proteins. Using newly developed methods of sequence analysis, in combination with protein modeling, we provide a framework for investigating the evolution and biological function of genes like the PSG. Evolutionary trees, based on C-terminal sequence, group PSG genes in a manner consistent with their genomic organization. Trees constructed using the N-terminal domain sequences are unreliable as an indicator of phylogeny because of non-neutral processes of sequence change. During duplication of the PSG genes, evolutionary pressures have resulted in a gradient of constrained change across each gene. The N-terminal domains show a nonrandom pattern of amino acid substitutions clustered in the immunoglobulin complementarity-determining region (CDR)-like regions, which appear to be important in the function of the protein. 相似文献
107.
108.
109.
Junko Komura Ikumi Tamai Mizuho Senmaru Tetsuya Terasaki Yoshimichi Sai Akira Tsuji 《Journal of neurochemistry》1996,67(1):330-335
Abstract: The characteristics of β-alanine transport at the blood-brain barrier were studied by using primary cultured bovine brain capillary endothelial cells. Kinetic analysis of the β-[3 H]alanine transport indicated that the transporter for β-alanine functions with Kt of 25.3 ± 2.5 µ M and J max of 6.90 ± 0.48 nmol/30 min/mg of protein in the brain capillary endothelial cells. β-[3 H]Alanine uptake is mediated by an active transporter, because metabolic inhibitors (2,4-dinitrophenol and NaN3 ) and low temperature reduced the uptake significantly. Furthermore, the uptake of β-[3 H]alanine required Na+ and Cl− in the external medium. Stoichiometric analysis of the transport demonstrated that two sodium ions and one chloride ion are associated with one β-alanine molecule. The Na+ and Cl− -dependent uptake of β-[3 H]alanine was stimulated by a valinomycin-induced inside-negative K+ -diffusion potential. β-Amino acids (β-alanine, taurine, and hypotaurine) inhibited strongly the uptake of β-[3 H]alanine, whereas α- and γ-amino acids had little or no inhibitory effect. In ATP-depleted cells, the uptake of β-[3 H]alanine was stimulated by preloading of β-alanine or taurine but not l -leucine. These results show that β-alanine is taken up by brain capillary endothelial cells, via the secondary active transport mechanism that is common to β-amino acids. 相似文献
110.
Effect of Hypoxia on Na+-K+-Cl− Cotransport in Cultured Brain Capillary Endothelial Cells of the Rat
Abstract: The effect of hypoxia on Na+,K+-ATPase and Na+-K+-Cl? cotransport activity in cultured rat brain capillary endothelial cells (RBECs) was investigated by measuring 86Rb+ uptake as a tracer for K+. RBECs expressed both Na+,K+-ATPase and Na+-K+-Cl? cotransport activity (4.6 and 5.5 nmol/mg of protein/min, respectively). Hypoxia (24 h) decreased cellular ATP content by 43.5% and reduced Na+,K+-ATPase activity by 38.9%, whereas it significantly increased Na+-K+-Cl? cotransport activity by 49.1% in RBECs. To clarify further the mechanism responsible for these observations, the effect of oligomycin-induced ATP depletion on these ion transport systems was examined. Exposure of RBECs to oligomycin led to a time-dependent decrease of cellular ATP content (by ~65%) along with a complete inhibition of Na+,K+-ATPase and a coordinated increase of Na+-K+-Cl? cotransport activity (up to 100% above control values). Oligomycin augmentation of Na+-K+-Cl? cotransport activity was not observed in the presence of 2-deoxy-d -glucose (a competitive inhibitor of glucose transport and glycolysis) or in the absence of glucose. These results strongly suggest that under hypoxic conditions when Na+,K+-ATPase activity is reduced, RBECs have the ability to increase K+ uptake through Na+-K+-Cl? cotransport. 相似文献