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91.
The yeast mitochondrial phosphate transport protein (PTP) has only 38% sequence similarity to the bovine heart protein, and it has recently been postulated to code for a mitochondrial import receptor. Since the reconstitutively active protein is not completely pure, it is important to demonstrate explicitly that the yeast gene codes for PTP. We have replaced Thr43 with Cys (T43C) and show that its unidirectional and pH gradient-dependent inorganic phosphate transport activity becomes highly sensitive to N-ethylmaleimide. This new PTP/T43C catalyzes less than 10% of the wild type transport activity (1 mM [Pi]e, pHe (6.80); 0 mM [Pi]i, pHi (8.07); 30 s [Pi] uptake) suggesting that Thr43 occupies an important position in the PTP. 相似文献
92.
Viscoelastic response of fibroblasts to tension transmitted through adherens junctions. 总被引:1,自引:0,他引:1 下载免费PDF全文
Cytoplasmic deformation was monitored by observing the displacements of 200-nm green fluorescent beads microinjected into the cytoplasm of Swiss 3T3 fibroblasts. We noted a novel protrusion of nonruffling cell margins that was accompanied by axial flow of beads and cytoplasmic vesicles as far as 50 microm behind the protruding plasma membrane. Fluorescent analog cytochemistry and immunofluorescence localization of F-actin, alpha-actinin, N-cadherin, and beta-catenin showed that the protruding margins of deforming cells were mechanically coupled to neighboring cells by adherens junctions. Observations suggested that protrusion resulted from passive linear deformation in response to tensile stress exerted by centripetal contraction of the neighboring cell. The time dependence of cytoplasmic strain calculated from the displacements of beads and vesicles was fit quantitatively by a Kelvin-Voight model for a viscoelastic solid with a mean limiting strain of 0.58 and a mean strain rate of 4.3 x 10(-3) s(-1). In rare instances, the deforming cell and its neighbor spontaneously became uncoupled, and recoil of the protruding margin was observed. The time dependence of strain during recoil also fit a Kelvin-Voight model with similar parameters, suggesting that the kinetics of deformation primarily reflect the mechanical properties of the deformed cell rather than the contractile properties of its neighbor. The existence of mechanical coupling between adjacent fibroblasts through adherens junctions and the viscoelastic responses of cells to tension transmitted directly from cell to cell are factors that must be taken into account to fully understand the role of fibroblasts in such biological processes as wound closure and extracellular matrix remodeling during tissue development. 相似文献
93.
94.
Prevention by uncouplers of lipophilic chelator inhibition at three sites of mitochondrial electron transport 总被引:2,自引:0,他引:2
D C Phelps H J Harmon F L Crane 《Biochemical and biophysical research communications》1974,59(4):1185-1191
Inhibition of coupled electron transport in mitochondrial cristae by hydrophobic chelators such as bathophenanthroline is partially prevented by uncoupling agents such as carbonylcyanide-m-chlorophenyl hydrazone. Reversal of inhibition is observed in all three sections of the electron transport chain where energy coupling occurs. Sites inhibited by hydrophilic bathophenanthroline sulfonate or orthophenanthroline do not show uncoupler relief of inhibition. We conclude that a buried non-heme iron or copper protein is closely associated with each energy coupling site. 相似文献
95.
The biosynthesis in vitro of chondroitin sulphate in neonatal rat epiphysial cartilage 总被引:5,自引:5,他引:0
1. A system is described, which was used to incubate neonatal rat epiphysial cartilage in vitro with [U-(14)C]glucose and [(35)S]sulphate. 2. The acid glycosaminoglycans of neonatal rat epiphyses were extracted and fractionated on cetylpyridinium chloride-cellulose columns. The major components were chondroitin 4-sulphate (65%), chondroitin 6-sulphate (15%), hyaluronic acid (4%) and keratan sulphate (2%). 3. The acid-soluble nucleotides and intermediates of glycosaminoglycan synthesis were separated on a Dowex 1 (formate) system. The tissue contents and cellular concentrations of these metabolites were determined. 4. The rates of synthesis of UDP-glucuronic acid and UDP-N-acetyl-hexosamine from [U-(14)C]glucose were found to be 0.79+/-0.16 and 3.2+/-0.08nmol/min per g wet wt. respectively. 5. The incorporation of [U-(14)C]glucose into the uronic acid and hexosamine moieties of the polymers was also measured and the turnover rates of the glycosaminoglycans were calculated. It was found that chondroitin sulphate was turning over in about 70h and hyaluronic acid in about 120h. 6. The relative rates of synthesis of the sulphated glycosaminoglycans were calculated from [(35)S]sulphate incorporation and were found to be in good agreement with those obtained from [U-(14)C]glucose labelling. 相似文献
96.
X-ray-diffraction patterns of hyaluronate fibres from a variety of sources were obtained. Sodium hyaluronate gives well-defined patterns which index on a hexagonal unit cell with dimensions a=1.17+/-nm and a fibre repeat-distance of 2.85+/-0.03nm. A further form of sodium hyaluronate is produced by annealing at 60 degrees C in 75% relative humidity. This stable state indexes on a hexagonal unit cell of unchanged fibre repeat-distance but with a=1.87nm. The chain conformation is a threefold helix. Analysis of these diffraction patterns led to two tentative structures for sodium hyaluronate, involving different packing of the polysaccharide chains. The significance of side-chain interaction is discussed. Hyaluronic acid produces an X-ray pattern different from that obtained with the sodium salt. The fibre repeat-distance is 1.96+/-0.02nm and the unit cell appears to be monoclinic. The chain conformation is a twofold helix and conformational change between free acid and monovalent salt is discussed. These findings, together with model-building experiments, are interpreted as indicating a highly ordered structure, and the physical properties of hyaluronate solutions with regard to molecular shape and polyelectrolyte behaviour are rationalized. 相似文献
97.
The neuropeptide vasopressin and its receptor V1aR are broadly implicated in social behavior and play a central role in several key aspects of male mating tactics in voles. In the prairie vole, a microsatellite in the cis-regulatory region of the gene encoding V1aR (avpr1a) provides a potential genetic basis for individual variation in neural phenotype and behavior; recent studies found that allele length predicts V1aR expression and male social attachment in the laboratory. Here, we explore the relationship between avpr1a microsatellite length, V1aR neural phenotype, and field measures of monogamy and fitness in male prairie voles. We found significant effects of allele length on V1aR expression in structures integral to pairbond formation. These effects did not, however, translate to differences in mating tactics or reproductive success. Together, these data suggest that, while length polymorphism in the avpr1a microsatellite influences neuronal phenotype, this variation does not contribute significantly to male reproductive success and field behavior. We propose that previously reported behavioral effects may be mediated primarily by sequence variation at this locus, for which allele length is an imperfect proxy. By combining genetic, neuronal and ecological approaches, these data provide novel insights into the contribution of genotype to natural diversity in brain and behavior. 相似文献
98.
99.
Joan F. Carroll Kimberly G. Fulda Ana L. Chiapa Mayra Rodriquez David R. Phelps Kathryn M. Cardarelli Jamboor K. Vishwanatha Roberto Cardarelli 《Obesity (Silver Spring, Md.)》2009,17(7):1420-1427
The purpose of this study was to determine whether racial/ethnic differences exist in the relationship between visceral adipose tissue (VAT) and selected inflammatory biomarkers. Subjects included 136 African‐American, 133 Hispanic, and 100 white men and women, aged ≥45. Waist circumference and BMI were measured using standard methods. Total VAT, and VAT and subcutaneous adipose tissue (SAT) at the L4L5 spinal level were measured using computed tomography. Interleukin‐6 (IL‐6), C‐reactive protein (CRP), and fibrinogen were measured from fasting blood samples. Results revealed that waist circumference and BMI were similar among groups but African Americans had significantly lower L4L5 VAT compared with Hispanics and whites. Despite lower VAT, African‐American men had similar concentrations of inflammatory biomarkers. On the other hand, African‐American women had higher CRP and IL‐6 than white women, and higher fibrinogen than both Hispanic and white women. After controlling for L4L5 VAT, L4L5 SAT, and age, African‐American women had higher concentrations of IL‐6 and fibrinogen. Stratified analyses for CRP indicated that L4L5 SAT was associated with CRP in African‐American and white women after controlling for L4L5 VAT and age, but that the reverse was not true. These data indicate that African Americans had lower VAT but similar or higher concentrations of inflammatory biomarkers. African‐American women consistently displayed greater inflammation compared with whites, even after controlling for VAT or SAT. 相似文献
100.
George TG Johnsamuel J Delfín DA Yakovich A Mukherjee M Phelps MA Dalton JT Sackett DL Kaiser M Brun R Werbovetz KA 《Bioorganic & medicinal chemistry》2006,14(16):5699-5710
N(1)-Phenyl-3,5-dinitro-N(4),N(4)-di-n-propylsulfanilamide (1) and N(1)-phenyl-3,5-dinitro-N(4),N(4)-di-n-butylsulfanilamide (2) show potent in vitro antimitotic activity against kinetoplastid parasites but display poor in vivo activity. Seventeen new dinitroaniline sulfonamide and eleven new benzamide analogs of these leads are reported here. Nine of the sulfonamides display in vitro IC(50) values under 500 nM against African trypanosomes, and the most active antikinetoplastid compounds also inhibit the in vitro assembly of purified leishmanial tubulin with potencies similar to that of 2. While several of the potent compounds are rapidly degraded by rat liver S9 fractions in vitro, N(1)-(3-hydroxy)phenyl-3,5-dinitro-N(4),N(4)-di-n-butylsulfanilamide (21) displays an IC(50) value of 260 nM against African trypanosomes in vitro and is more stable than 2 in the in vitro metabolism assay. 相似文献