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31.
Direct mass determination of purified SV40 minichromosomes was obtained by scanning transmission electron microscopy. Twenty to thirty percent of the minichromosomes were found with an Mr of 6.9±0.4×106. The rest of the molecules formed a spread Mr distribution ranging from 7.3×106 to 9.5×106 due possibly to different contents of the virus-coded proteins, mainly VP1. The apparent mass histogram of individual SV40 nucleosomes presents three maxima at Mr 2.1×105, 2.6×105 and 3.1×105 that could correspond to partially unravelled nucleosomes, complete nucleosomes and complete nucleosomes with the addition of VP1. Beaded structures with a higher mass were also measured; some were found at either side of the open nucleosome-free region. 相似文献
32.
Isolation of cDNA clones and complete amino acid sequence of human erythrocyte glycophorin C 总被引:7,自引:0,他引:7
Y Colin C Rahuel J London P H Roméo L d'Auriol F Galibert J P Cartron 《The Journal of biological chemistry》1986,261(1):229-233
Two cDNA clones for glycophorin C, a transmembrane glycoprotein of the human erythrocyte which carries the blood group Gerbich antigens, have been isolated from a human reticulocyte cDNA library. The clones were identified with a mixture of 32 oligonucleotide probes (14-mer) which have been synthetized according to the amino acid sequence Asp-Pro-Gly-Met-Ala present in the N-terminal tryptic peptide of the molecule. The primary structure of glycophorin C deduced from the nucleotide sequence of the 460 base-pair insert of the pGCW5 clone indicates that the complete protein is a single polypeptide chain of 128 amino acids clearly organized in three distinct domains. The N-terminal part (residues 1-57, approximately) which is N- and O-glycosylated is connected to a hydrophilic C-terminal domain (residues 82-128, approximately) containing 4 tyrosine residues by a hydrophobic stretch of nonpolar amino acids (residues 58-81, approximately) probably interacting with the membrane lipids and permitting the whole molecule to span the lipid bilayer. Northern blot analysis using a 265-base-pair restriction fragment obtained by DdeI digestion of the inserted DNA shows that the glycophorin C mRNA from human erythroblasts is approximately 1.4 kilobases long and is present in the human fetal liver and the human K562 and HEL cell lines which exhibit erythroid features. The glycophorin C mRNA, however, is absent from adult liver and lymphocytes, indicating that this protein represents a new erythrocyte-specific probe which might be useful to study erythroid differentiation. 相似文献
33.
Characteristics of the β-Adrenoreceptor from Neuronal and Glial Cells in Primary Cultures of Rat Brain 总被引:1,自引:1,他引:0
Stephen P. Baker Colin Sumners Joseph Pitha† Mohan K. Raizada 《Journal of neurochemistry》1986,47(4):1318-1326
The cellular characteristics of the beta-adrenoreceptor in glial and neuronal cells from the newborn rat brain were determined by (-)-[125I]iodocyanopindolol binding. In membranes from both cell types, the binding was saturable and from competition assays the potency series of (-)-isoproterenol greater than (-)-epinephrine = (-)-norepinephrine greater than (+)-isoproterenol was observed. 5'-Guanylyl-imidodiphosphate reduced the affinity of (-)-isoproterenol for the beta-adrenoreceptor from glial cells but had no effect on agonist affinity in neuronal cells. Chronic treatment of both cell types with (-)-isoproterenol reduced the receptor content and the capacity of the agonist to increase the cellular cyclic AMP content. However, the receptor recovery after chronic agonist treatment was faster in glial cells (72 h) than neuronal cells (120 h) and was blocked by cycloheximide. Treatment of both types with the irreversible beta-blocker bromoacetylalprenololmentane (2 microM) reduced the receptor content by 78% but no receptor recovery was observed for 120 h after the initial receptor loss. The data indicated that the majority of beta-adrenoreceptors in both cell types are the beta-1 subtype, but show some differences in receptor-agonist interactions. Furthermore, these CNS cells may be useful models for regulatory studies on the beta-adrenoreceptor. 相似文献
34.
α1 -Adrenergic Receptor-Mediated Downregulation of Angiotensin II Receptors in Neuronal Cultures 总被引:1,自引:0,他引:1
Previous evidence has suggested that brain catecholamine levels are important in the regulation of central angiotensin II receptors. In the present study, the effects of norepinephrine and 3,4-dihydroxyphenylethylamine (dopamine) on angiotensin II receptor regulation in neuronal cultures from rat hypothalamus and brainstem have been examined. Both catecholamines elicit significant decreases in [125I]angiotensin II-specific binding to neuronal cultures prepared from normotensive rats, effects that are dose dependent and that are maximal within 4-8 h of preincubation. Saturation and Scatchard analyses revealed that the norepinephrine-induced decrease in the binding is due to a decrease in the number of angiotensin II receptors in neuronal cultures, with little effect on the receptor affinity. Norepinephrine has no significant actions on [125I]angiotensin II binding in cultures prepared from spontaneously hypertensive rats. The downregulation of angiotensin II receptors by norepinephrine or dopamine is blocked by alpha 1-adrenergic and not by other adrenergic antagonists, a result suggesting that this effect is initiated at the cell surface involving alpha 1-adrenergic receptors. This is further supported by our data indicating a parallel downregulation of specific alpha 1-adrenergic receptors elicited by norepinephrine. In summary, these results show that norepinephrine and dopamine are able to alter the regulation of neuronal angiotensin II receptors by acting at alpha 1-adrenergic receptors, which is a novel finding. 相似文献
35.
I. Colin Prentice 《Plant Ecology》1986,67(2):131-141
Vegetation responses to climatic change can be studied retrospectively by utilizing the Quaternary fossil record. There has been controversy over the extent to which major changes in vegetation patterns at the continental scale lag behind the climatic changes that drive them, and to what extent vegetation can ever be said to be in equilibrium with climate. The equilibrium question has no single answer. The predominant mode of vegetation response to climatic change depends on the space and time frame and resolution of the data set in which the response is observed.Vegetation (as observed on particular space and time scales) can be in dynamic equilibrium with climate if its response time is sufficiently fast in relation to the rate of climatic change to which it is observed to be responding. Several processes can be involved in the response: successional, migrational, edaphic, and evolutionary. Successional response times can be deduced from forest succession models. The other processes are less well understood and different ideas exist concerning their rates. According to one hypothesis, migrational lags caused delays of thousands of years in the postglacial dynamics of forest composition. The alternative hypothesis explains these changes as dynamic equilibrium responses to changes in climatic seasonality and climatic anomaly patterns. Neither hypothesis need be universally true; gradient analysis and forest succession models are among the techniques that can be used in inferential tests of these hypotheses for particular space-time regions.Dynamic equilibrium may often be a reasonable approximation for the responses of the broadest continental-scale forest patterns to orbitally induced climatic changes. But as spatial and temporal frames of observation are diminished and resolution increased, biotic processes must eventually come to dominate. At sufficiently fine scales the main observable phenomena are successional responses to natural disturbance events. The late-Quaternary record of vegetation change allows a choice of observation scales and thus provides a continuum of possibilities for study, ranging from long-term dynamic bioclimatology to more conventional vegetation dynamics.I thank Margaret Davis, Honor Prentice, Jim Ritchie, Al Solomon, Geoff Spaulding and Tom Webb for their reviews of earlier drafts. Research supported by a US Department of Energy, Carbon Dioxide Research Division, grant to Brown University and a Swedish Natural Science Research Council grant to the project SlsSimulation of Natural Forest Dynamics.I thank Margaret Davis, Honor Prentice, Jim Ritchie, Al Solomon, Geoff Spaulding and Tom Webb for their reviews of earlier drafts. Research supported by a US Department of Energy, Carbon Dioxide Research Division, grant to Brown University and a Swedish Natural Science Research Council grant to the project SlsSimulation of Natural Forest Dynamics. 相似文献
36.
Plant Molecular Biology - The α-Amy1 and α-Amy2 genes of wheat produce distinct subsets of α-amylase isozymes which show different patterns of expression in wheat aleurone cells and... 相似文献
37.
Dr. Sasha Malamed Jean A. Gibney Kathryn E. Loesser Colin G. Scanes 《Cell and tissue research》1985,239(1):87-91
Summary The ultrastructure of the somatotrophs of the caudal pituitary of the domestic fowl was studied quantitatively. Two age groups of male chickens were compared: 4–6 weeks and 24–30 weeks post-hatching. With age, somatotrophs decreased from about 40% to about 30% of the pituitary cell population. Their volume density decreased similarly. Mean volume of a somatotroph was the same in young and adult animals. Because the granule volume density of the somatotrophs was unchanged, but the somatotroph volume density of the gland declined, the granule volume density of the caudal pituitary gland dropped in parallel with that of the somatotrophs. Thus the volume of the gland comprised of somatotroph granules fell about 32%: from 6.57% to 4.45%. This lowered pool of stored hormone may be linked to the lowered circulating levels of growth hormone found in older animals by other investigators.The granule volume density of the somatotrophs was unchanged but the numerical density approximately doubled; thus the mean granule size decreased by 47% with age. The relationship of the size reduction of the granules to the lowered plasma growth hormone levels is not understood at present.Supported in part by Hatch and State funds from the New Jersey State Agricultural Experimental Station and NSF grants PCM 8,0227,27 and PCM 8,302197. 相似文献
38.
Anil K. Sinha V. Ann Linscombe B. Bhaskar Gollapudi George C. Jersey Colin N. Park 《Cell biology and toxicology》1985,1(4):333-342
Sister chromatid exchanges (SCE) were analyzed in peripheral blood lymphocytes from a select group of 71 healthy men, 56 nonsmokers and 15 cigarette smokers. In addition to estimating baseline SCE, data were examined to seek relationships of SCE frequencies to age and smoking. The baseline value of 7.53 SCE per cell from the 56 nonsmokers was within the range (5.60 to 9.10 SCE/cell) reported for other human populations. No relationship was found between the mean SCE frequency per cell and age. However, a significant increase in the SCE mean value was observed in smokers as compared to nonsmokers. The results of this study are compared with those of other reports on SCE effects of age and smoking.Abbreviations BUdR
5-bromo,2-deoxyuridine
- SCE
sister chromatid exchange 相似文献
39.
Respiratory chain phosphorylation has been investigated in the methylotrophic bacterium Methylophilus methylotrophus following the addition of oxidisable substrates to aerobic, whole cell suspensions. Initial-rate experiments showed that ATP synthesis occurred at the overall expense of AMP and inorganic phosphate via the sequential action of the ATP phosphohydrolase and adenylate kinase; some of the nascent ATP was rapidly used to synthesis nonadenine nucleoside triphosphates. After being corrected for ATP turnover, Pi/O quotients of 0.46 to 0.54, 0.77 and 1.37 nmol/ng-atom O were obtained for the oxidation of methanol dehydrogenase-linked substrates (methanol, ethanol and acetaldehyde), duroquinol and formate (NAD+-linked) respectively. These values were proportional to the H+/O and/or K+/O quotients exhibited by these substrates, and yielded an average H+/ATP (H+/Pi) quotient of 4.2 ng-ion H+/nmol. Steady-state experiments showed that the extent of cellular energisation varied with the respiration rate but was always in the order methanol > duroquinol > acetaldehyde, thus indicating that under these longer-term conditions methanol was completely oxidised to yield PQQH2 and 2NAD(P)H. These results are discussed in terms of the various reactions which lead to the generation or utilisation of the protonmotive force in this organism.Abbreviations FCCP
carbonylcyanide p-trifluoromethyxyphenyl-hydrazone
-
bulk phase, transmembrane electrochemical potential difference of protons (
)
- pH
bulk phase, transmembrane pH difference (pHin–pHout)
-
bulk phase, transmembrane electrical potential difference (in - out)
- [P]
concentration of anhydride phosphate bonds in adenine nucleotides (2[ATP]+[ADP])
- FPLC
fast protein liquid chromatography
- PQQ
pyrroloquinoline quinone
- Gp
phosphorylation potential 相似文献
40.
Colin J. Henderson Martin J. Hulme R. John Aitken 《Molecular reproduction and development》1987,16(4):323-341
Zonae pellucidae (ZP) were isolated from 1,500 porcine ovaries and heat solubilized to generate approximately 15 mg ZP glycoprotein. Analysis of this material by isoelectric focusing, one-dimensional electrophoresis, and gas chromatography indicated the presence of a major glycoprotein species that exhibited considerable microheterogeneity with respect to its charge (pI 7.5–3.5) and molecular mass (45–85 kDa) and that contained 39.6% carbohydrate, predominantly N-acetylglucosamine. Chemical deglycosylation of porcine ZP using trifluoromethanesulphonic acid (TFMS) resulted in the production of five discrete protein bands on one-dimensional sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS/PAGE) with molecular masses of 66, 52, 36, 32, and 16 kDa. Antisera raised in rabbits and marmosets to ZP and/or deglycosylated ZP (DGZP) were used in immunoblotting experiments to demonstrate the retention of immunogenicity by DGZP and the cross-reactivity of the antisera with their heterologous antigen. These studies indicated that antisera that were capable of inhibiting the fertility of primates in vivo and the penetration of the human ZP in vitro reacted preferentially with 3 of the 5 products of deglycosylation, with molecular masses of 66, 52, and 36 kDa. Anti-DGZP antibodies were also shown to interact with intact porcine and human ZP and, with the latter, to block the ability of human spermatozoa to both bind to and penetrate this structure. 相似文献