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151.
Charybdotoxin block of Shaker K+ channels suggests that different types of K+ channels share common structural features 总被引:9,自引:0,他引:9
Charybdotoxin (CTX), a 37 amino acid protein isolated from the venom of L. quinquestriatus, is a high-affinity blocker of various Ca2(+)-activated K+ channels. CTX also blocks Drosophila Shaker (Sh) clone H4 transient K+ currents expressed in Xenopus oocytes with similar affinity (Kd = 3.6 nM). CTX blocks both the open and the closed states of Sh channels with no apparent change in gating behavior. In addition, the block is enhanced as the ionic strength is lowered. These properties are identical to those of CTX block of Ca(+)-activated K+ channels, and these results suggest that the external pore openings of these two functionally dissimilar K+ channels may share common structural features. 相似文献
152.
Development and characterization of continuous avian cell lines depleted of mitochondrial DNA 总被引:6,自引:0,他引:6
Réjean Morais Paul Desjardins Chanta Turmel Karen Zinkewich-Péotti 《In vitro cellular & developmental biology. Plant》1988,24(7):649-658
Summary Populations of quail and chicken cells were treated with ethidium bromide, an inhibitor of mitochondrial DNA replication.
After long-term exposure to the drug, the cell populations were transferred to ethidium bromide (EtdBr)-free medium, and cloned.
Clones HCF7 (quail) and DUS-3 (chicken) were propagated for more than a year, and then characterized. Analysis of total cellular
DNA extracted from these cells revealed no characteristic mitochondrial DNA molecule by Southern blot hybridization of HindIII-
or AvaI-digested total cellular DNA probed with cloned mitochondrial DNA fragments. Reconstruction experiments, where a small
number of parental cells was mixed with HCF7 cells and DUS-3 cells before extraction of total cellular DNA, further strengthen
the notion that the drug-treated cells are devoid of mitochondrial DNA molecules. The cell populations were found to proliferate
at a moderately reduced growth rate as compared to their respective parents, to be auxotrophic for uridine, and to be stably
resistant to the growth inhibitory effect of EtdBr and chloramphenicol. At the ultrastructural level, mitochondria were considerably
enlarged and there was a severe reduction in the number of cristae within the organelles and loss of cristae orientation.
Morphometric analysis revealed a fourfold increase of the mitochondrial profile area along with a twofold decrease of the
numerical mitochondrial profiles. Analysis of biochemical parameters indicated that the cells grew with mitochondria devoid
of a functional respiratory chain. The activity of the mitochondrial enzyme dihydroorotate dehydrogenase was decreased by
95% and presumably accounted for uridine auxotrophy.
This work was supported by a grant from the Medical Research Council of Canada. 相似文献
153.
154.
Charles R. Ill Tammy Brehm Bjorn K. Lydersen Rachel Hernandez Karen G. Burnett 《In vitro cellular & developmental biology. Plant》1988,24(5):413-419
Summary Studies with Human x Human (HxH), Human x Mouse (HxM), and Mouse x Mouse (MxM) hybridomas have enabled us to define specific
factors that affect hybridoma growth in a species-specific manner. Three transferrins and three lipophilic iron chelates have
been tested for their ability to support hybridoma proliferation and antibody production. The results of these studies demonstrate
that HxH hybridomas do not respond to bovine transferrin a+ concentrations up to 100 μg/ml and are approximately 100-fold less responsive to mouse transferrin than to human transferrin.
HxM and MxM hybridomas respond equally to human or mouse transferrin but are 100-fold less sensitive to bovine transferrin.
An antibody to the human transferrin receptor inhibited the growth-promoting activity of human or mouse transferrin on HxH
hybridomas but was ineffective on HxM hybridomas. This semonstrated the functionality of the human transferrin receptor in
HxH hybridomas and that human, mouse, and bovine transferrin were interacting through the mouse transferrin receptor in HxM
hybridomas. HxH and HxM hybridomas respond similarly to three different iron chelates exhibiting 80 to 110% of the growth
response to human transferrin. MxM hybridomas fail to respond to the iron chelates at similar concentrations, suggesting that
the human genome present in the other hybridoma species confers a unique ability for utilizing iron when delivered in this
form. 相似文献
155.
J. M. White D. D. Jones D. Huang J. J. Gauthier 《Journal of industrial microbiology & biotechnology》1988,3(5):263-272
Summary A cyanide-degrading pseudomonad was isolated by selective enrichment in a chemostat inoculated with coke-plant activated sludge and maintained at a dilution rate of 0.042/h for 60 days with a feed of 10 mg/l cyanide. The isolate, a facultative methylotroph capable of growth on methanol and methylamine, degraded cyanide to formate and ammonia; it could utilize the released ammonia as a nitrogen source but did not further metabolize formate under the experimental conditions employed. Both cyanide-degrading enzyme activity and respiratory resistance to cyanide were inducible and were enhanced by repeated exposure to the compound. Cell-free extracts stoichiometrically converted cyanide to formate and ammonia in a reaction that did not require oxygen. Enzyme activity, lost upon dialysis, was restored by less than equimolar ratios of NAD(P)H or ascorbate to cyanide, indicating that the reductants did not function directly as co-enzymes. 相似文献
156.
Keith C. White Patricia C. Babbitt Douglas D. Buechter George L. Kenyon 《Journal of Protein Chemistry》1992,11(5):489-494
The importance of creatine kinase (E.C. 2.7.3.2) in endocrine tissues has been generally overlooked. Using a specific radiometric assay, we have demonstrated the existence of CK in the Brockmann body (principal islet) of the Coho salmon. We have purified this protein from insular tissue and concurrently purified CK from brain and muscle of the salmon. Purification characteristics, immunological cross-reactivity, and N-terminal sequence analysis have demonstrated that the predominant cytosolic CK from the Brockmann body is indistinguishable from the BB (brain) isoenzyme. Immunocytochemical studies indicated that the enzyme resides in the endocrine parenchyma. Phosphocreatine may serve as a reservoir of energy in the islet and augment its capacity to secrete hormones. The induction of CK-BB in the islet by other hormones could influence the secretion of insular hormones. Interorgan flux of the substrate creatine may be an undescribed mechanism of physiological regulation. 相似文献
157.
Summary Aberrations of photoreceptor ultrastructure resulting from carotenoid/retinoid (vitamin A) deprivation were studied in the retina of Manduca sexta. The syndrome of chromophore deficiency included hypertrophy of smooth endoplasmic reticulum, variable dilation of rhabdomeric microvilli, the insertion of endomembrane fingers into such enlarged microvilli, and the formation of rhabdomeric vacuoles, intracellular compartments containing microvilli similar to those of the rhabdomere. Retinas were processed either with conventional procedures employing preliminary aldehyde fixation followed by heavy metal postfixation, or by fixation and incubation in unbuffered OsO4. The latter method deposits osmium throughout the endomembrane system, within the rhabdomeric vacuoles, and in the extracellular space of the rhabdom. However, the intravillous fingers were rarely impregnated with osmium, despite their continuity with densely stained cisternae of the smooth endoplasmic reticulum. We suggest that the insertion of endomembrane fingers into dilated microvilli results from a cytoskeleton-mediated link between cisternae of the smooth endoplasmic reticulum and the rhabdomeric membrane, an association that may be important in the turnover of photoreceptor membrane. We interpret endomembrane hypertrophy and development of rhabdomeric vacuoles as symptoms of disturbance in the pathway leading to the assembly of the rhabdomere resulting from reduced synthesis of visual pigment. 相似文献
158.
Besides Clostridium thermoaceticum and C. formicoaceticum other resting acetogenic clostridia such as C. aceticum and C. thermoautotrophicum and to a lesser extent non-clostridial acetogens such as Butyribacterium methylotrophicum and Eubacterium limosum were able to reduce propionate to propanol at the expense of carbon monoxide or formate. Methylviologen usually increased the reduction rate. Ten M molybdate in the growth medium decreased this capability for C. thermoaceticum but increased it or had no effect for the other organisms. Ten M tungstate in the growth medium increased the aldehyde oxidoreductase activity in all organisms. Crude extracts of C. thermoaceticum cells grown in the presence of 10 M or 1 mM molybdate showed by ELISA the same or even a 4 fold concentration of aldehyde oxidoreductase in the latter case. However, the enzymic activity was very low in both cases. Omission of dithionite in the growth medium diminished the antigen by a factor of about 8. The immunological distance between the enzyme from C. thermoaceticum and C. thermoautotrophicum was rather low but very large to C. formicoaceticum and undeterminably large to the other organisms.Abbreviations Ald-DH
aldehyde dehydrogenase
- AOR
aldehyde oxidoreductase
- CO-DH
carbon-monoxide dehydrogenase
- ELI-SA
enzyme-linked immunosorbent assay
- FDH
formate dehydrogenase
- MV
methylviologen
- V++
oxidised
- V+.
reduced viologen 相似文献
159.
Sasha N. Zill S. Faith Frazier John Lankenau Karen Jepson-Innes 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1992,170(6):761-772
Summary The use of the locust (Schistocerca americana) hindleg in postural control was examined in animals that stood on a repeatedly swayed vertical substrate. Myograms were recorded from leg muscles and the angle of the femoro-tibial joint was monitored photographically. Two discrete strategies were observed,; in compensatory reactions the hindleg was held in place, while in flexion reactions, the leg was moved, most often to complete flexion of the femoro-tibial joint. Tightly coupled, rhythmic bursting occurred in the flexor and levator muscles of the leg during compensatory reactions. Bursting was initiated repeatedly when the substrate was being pulled away from the animal. Bursting was correlated with subsequent decreases in the rate of change of the femorotibial joint angle. Compensatory and flexion reactions occurred preferentially in different ranges of joint angles: most often, compensatory reactions occurred in the midrange, while flexion reactions were elicited in the extremes of joint angle. These differences may be due to the mechanical advantages of the tibial muscles and the leg may be moved to full flexion because of a locking mechanism of the flexor muscle tendon. These reactions are compared with known reflexes of hindleg proprioceptors and contrasted with similar responses of vertebrates. 相似文献
160.
Phyllis W. Speisere Maria I. New Grace M. Tannin Donald Pickering Soo Young Yang Perrin C. White 《Human genetics》1992,88(6):647-648
Summary An A-to-G transition in the second intron was the sole mutation detected in four Yupik Eskimo patients with salt-wasting congenital adrenal hyperplasia due to steroid 21-hydroxylase deficiency. Allele-specific hybridization should be an efficient means of performing prenatal diagnosis of the disease in this highly inbred population. 相似文献