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11.
Brain tubulin was labeled in vitro by post-translational incorporation of [14C]-tyrosine or in vivo by intra-cranial injection of [3H]-leucine. The labeled protein was purified by ion-exchange chromatography. After incubating at 37 degrees C with a microsomal membrane preparation from rat brain, part of the labeled soluble tubulin became sedimentable at high-speed centrifugation. This was independent of the native configuration of tubulin, the state of tyrosination of the COOH-terminus, or the presence of 100 microM colchicine in the mixture. In addition, the double-labeled tubulin-colchicine complex obtained from the binding of [3H]-colchicine to [14C]-tyrosinated tubulin, bound to the membrane preparation to the same extent as [14C]-tyrosinated tubulin. The data show that either tubulin or the complex resulting from its binding to colchicine distributed between the soluble and the membrane fractions when mixed at 37 degrees C with a microsome preparation. Seemingly, the site for colchicine binding to tubulin needs not to be free for the protein-membrane association. 相似文献
12.
13.
Eugene L. Passer Rudean H. Leinaeng Lennel W. Birmingham Hector Cruz Chantale Dupervil Elizabeth J. Persaud Emil P. Dolensek 《Zoo biology》1989,8(4):357-365
This research determined the relationships between blood lead level and zinc protoporphyrin (ZnPROTO), protoporphyrin IX (PROTO), and free erythrocyte protoporphyrin (FEP) levels in a group of 18 ring teal ducks. Blood samples were drawn from two groups of ring teal ducks as part of the routine health maintenance program of the New York Zoological Park. One group of six teals had been exposed to what is believed to be lead-contaminated dust, while the second group of twelve teals was unexposed. Blood samples were analyzed for lead by flameless atomic absorption and for protoporphyrins by fluorescence. Blood lead level and log blood lead level had positive correlations with each of the protoporphyrins: the logarithmic correlations were better than the nonlogarithmic correlations, and PROTO correlated better than ZnPROTO. With one exception, PROTO levels were higher than ZnPROTO levels. The results suggest that PROTO, FEP, or ZnPROTO could serve as a biological indicator of lead poisoning in ring teal ducks. 相似文献
14.
Hector M. Alvarez Frank Mayer Dirk Fabritius A. Steinbüchel 《Archives of microbiology》1996,165(6):377-386
An oleaginous hydrocarbon-degrading Rhodococcus opacus strain (PD630) was isolated from a soil sample. The cells were able to grow on a variety of substrates and to produce large
amounts of three different types of intracellular inclusions during growth on alkanes, phenylalkanes, or non-hydrocarbon substrates.
Electron microscopy revealed large numbers of electron-transparent inclusions with a sphere-like structure. In addition, electron-dense
inclusions representing polyphosphate and electron-transparent inclusions with an elongated disc-shaped morphology occurred
in small amounts. The electron-transparent inclusions of alkane- or gluconate-grown cells were composed of neutral lipids
(98%, w/w), phospholipids (1.2%, w/w), and protein (0.8%, w/w). The major component of the cellular inclusions was triacylglycerols;
minor amounts of diacylglycerols and probably also some free fatty acids were also present. Free fatty acids and/or fatty
acids in acylglycerols in cells of R. opacus amounted up to 76 or 87% of the cellular dry weight in gluconate- or olive-oil-grown cells, respectively. The fatty acid composition
of the inclusions depended on the substrate used for cultivation. In cells cultivated on n-alkanes, the composition of the
fatty acids was related to the substrate, and intermediates of the β-oxidation pathway, such as hexadecanoic or pentadecanoic
acid, were among the acylglycerols. Hexadecanoic acid was also the major fatty acid (up 36% of total fatty acids) occurring
in the lipid inclusions of gluconate-grown cells. This indicated that strain PD630 utilized β-oxidation and de novo fatty
acid biosynthesis for the synthesis of storage lipids. Inclusions isolated from phenyldecane-grown cells contained mainly
the non-modified substrate and phenylalkanoic acids derived from the hydrocarbon oxidation, such as phenyldecanoic acid, phenyloctanoic
acid, and phenylhexanoic acid, and approximately 5% (w/w) of diacylglycerols. The lipid inclusions seemed to have definite
structures, probably with membranes at their surfaces, which allow them to maintain their shape, and with some associated
proteins, probably involved in the inclusion formation.
Received: 22 December 1995 / Accepted: 12 March 1996 相似文献
15.
Batdorj Enkhjin AlOgayil Najla Zhuang Qinwei Kim-wee Galvez Jose Hector Bauermeister Klara Nagata Kei Kimura Tohru Ward Monika A. Taketo Teruko Bourque Guillaume Naumova Anna K. 《Mammalian genome》2023,34(1):44-55
Mammalian Genome - Several lines of evidence suggest that the presence of the Y chromosome influences DNA methylation of autosomal loci. To better understand the impact of the Y chromosome on... 相似文献
16.
Solubilized components of the vitamin D3-25-hydroxylase, isolated from intact rat liver microsomes known to catalyze the C-25 oxidation of vitamin D3in vitro, have been separated into two submicrosomal fractions enriched in detergent-solubilized NADPH-cytochrome c reductase and cytochrome P-450 or P-448. The P-450 hemoprotein-containing fraction was obtained by solubilization with cholic acid followed by treatment with the nonionic detergent, Emulgen 911, yielding a final preparation with a specific content of 7.25 nmol/mg microsomal protein. The reduced triphosphopyridine nucleotide-dependent cytochrome P-450 reductase activity, as detected by its ability to reduce the artificial electron acceptor, cytochrome c, was isolated free of cytochromes b5 or P-450 by solubilization with deoxycholate and chromatography on DEAE-cellulose. The reductase component was found to exhibit kinetic properties with Michaelis constants: Km(NADPH) = 3.14 μM, Km(NADH) = 31.25 μM, and Km(cyt c) = 12.34 μM. The NADPH-cytochrome c reductase activity was sensitive to NADPH-reversible inhibition by NADP, but not rotenone or cyanide. When the isolated components were incubated in the presence of an NADPH-generating system and carbon monoxide under anaerobic conditions, enzymatic reduction of the P-450 hemoprotein was measured by the appearance of characteristic absorbances at 420 and 450 nm of the reduced carbon monoxide vs. reduced difference spectrum. Furthermore, when the soluble submicrosomal components were reconstituted with excess reduced triphosphopyridine nucleotide, 3H-labeled vitamin D3, and soluble cytosolic supernatant, full vitamin D3-25-hydroxylase activity was restored at rates of up to 7.68 pmol/h/mg protein, with an apparent turnover number of cytochrome P-450 of 1.16 to 1.20 under conditions where the concentrations of the hemoprotein were rate limiting for net product formation. These results strongly support the hypothesis that the rat liver microsomal mixed-function oxidase, vitamin D3-25-hydroxylase, consists of at least two membrane-bound protein components, NADPH-cytochrome c reductase and a cytochrome P-450 terminal oxidase, for the catalytic conversion of vitamin D3 to 25-hydroxyvitamin D3. 相似文献
17.
Summary The apical and basal-lateral plasma membranes of toad bladder epithelium were radio-iodinated with the glucose-glucose oxidase-lactoperoxidase system. The covalently bound radio-iodine was used as a marker during subcellular fractionation and membrane isolation. Homogenization conditions that ensured rupture of more than 80% of the cells without substantial nuclear damage were defined by Nomarski optics. The nuclei were separated by differential centrifugation and the apical and basal-lateral components were resolved by differential and sucrose density gradient centrifugation. The apical components yielded two radioactive bands that were identified as glycocalyx and plasma membrane labeled with125I. The basal-lateral components yielded a heterodisperse pattern made up of at least 3 radioactive bands, but the bulk of the activity of ouabain-sensitive ATPase comigrated with only one of these bands. The mitochondia, identified by assays for cytochrome oxidase and NADH cytochromec reductase activities, were separated from the radio-iodine labeled components by centrifugation in sucrose density gradients under isokinetic conditions. The labeled glycocalyx and the slowly migrating components of basal-lateral labeling were separated from the radio-iodinated membranes by centrifugation at 100,000 × g × 1 hr after removal of the mitochondria by the isokinetic method. The labeled membranes were then subjected to ultracentrifugation in sucrose density gradients under isopycnic conditions; the basal-lateral membranes containing ouabain-sensitive ATP-ase were well resolved from the apical membranes by this method. These results provide a relatively rapid method of attaining partial purification of the apical and basal-lateral plasma membranes of toad bladder epithelium. 相似文献
18.
Paul A. del Giorgio Alicia L. Vinocur Ruben J. Lombardo Hector G. Tell 《Hydrobiologia》1991,224(3):129-154
The phytoplankton of the River Lujan (Buenos Aires, Argentina) was studied for a period of 18 months, together with physical and chemical variables, in relation to a pollution gradient. 167 taxa were recorded within a seasonal succession characterized by dominance of diatoms with a brief summer green algae facies. A combination of several biotic indices and multivariate analysis was employed to assess the impact of pollution on the phytoplankton community. The biotic indices used were species diversity and richness, algal quotients (green algae/diatom ratio, Centrales/Pennales ratio) and the SD succession rate index. Multivariate procedures included cluster analysis and ordination by PCA of both species and samples, stepwise discriminant analysis and multiple discriminant analysis of variance (MANOVA). Results indicate that community dynamism is attenuated at the more polluted sites, concomitant with an increased predominance of a broad-tolerance algal assemblage, co-dominated by Cyclotella meneghiniana and Nitzschia stagnorum. The changes in the community structure and dynamics described herein involved alterations in the distribution and relative proportions of the algae, rather than modifications in the basic species composition. These changes may not be readily detectable by methods which over-simplify the ecological information, such as systems of indicator species and biotic indices, designed to assess the degree of pollution. The suitability of multivariate analysis and biotic indices in river phytoplankton studies is further discussed. 相似文献
19.
Postnatal Changes in the Activity Ratio of Specific and Nonspecific Cholinesterases from Neuronal Perikarya 总被引:3,自引:2,他引:1
Enrique L. M. Ochoa Alicia Brusco Victor P. Idoyaga-Vargas† Hector Carminatti† 《Journal of neurochemistry》1982,38(5):1236-1240
Abstract: A soluble fraction from rat brain neuronal perikarya was shown to contain both the specific and nonspecific forms of the enzyme acetylcholines-terase (EC's 3.1.1.7. and 3.1.1.8., respectively). The ratio of the enzyme activities varied along the course of brain development: the nonspecific form being predominant from 1 to 15 days of age and the specific one showing the pattern of rising activity from day 15 onward. We suggest a possible relationship between this changing in cholinesterase activities and the establishment of synapses within the rat cerebral cortex. 相似文献
20.
George A. Wudiri Suzanne M. Pritchard Hong Li Jin Liu Hector C. Aguilar Stacey D. Gilk Anthony V. Nicola 《Journal of virology》2014,88(23):13918-13922
Herpes simplex virus 1 (HSV-1) required cholesterol or desmosterol for virion-induced membrane fusion. HSV successfully entered DHCR24−/− cells, which lack a desmosterol-to-cholesterol conversion enzyme, indicating that entry can occur independently of cholesterol. Depletion of desmosterol from these cells resulted in diminished HSV-1 entry, suggesting a general sterol requirement for HSV-1 entry and that desmosterol can operate in virus entry. Cholesterol functioned more effectively than desmosterol, suggesting that the hydrocarbon tail of cholesterol influences viral entry. 相似文献