共查询到20条相似文献,搜索用时 31 毫秒
1.
G G Foster 《Developmental biology》1973,32(2):282-296
Temperature-shift experiments were performed on five Notch-locus genotypes with temperature-sensitive phenotypes. The results show that temperature-sensitive periods (TSPs) for lethality may occur at any developmental stage: (1) ;Dp51b7 having a short embryonic TSP for lethality, (2) having a second-instar TSP for lethality, and (3) with a long, possibly polyphasic, TSP, beginning in the embryonic stage and ending in the pupal stage. On the other hand, TSPs for adult morphological phenotypes appear to be restricted to the third larval instar: (1) having third-instar TSPs for wing vein gapping and ocellar bristle loss, and (2) having third-instar TSPs for eye facet disarray, wing notching, bristle number variation, and fusion of tarsal segments. The significance of these results is discussed in terms of the role of the Notch locus in development. 相似文献
2.
Transfer of N, N'-diacetylchitobiose from dolichyl diphosphate into a gray matter membrane glycoprotein 总被引:1,自引:0,他引:1
Gray matter and white matter membranes catalyze the transfer of label from UDP-N-acetyl-[] glucosamine into N-acetyl[]glucosaminyl-pyrophosphoryl-dolichol, N,N′-diacetyl []chitobiosyl-pyrophosphoryl-dolichol, and N-acetyl[]glucosamine-labeled glycoprotein. Gel filtration of the Pronase digests of gray matter N-acetyl[]glucosamine-labeled glycoprotein reveals two N-acetyl[]glucosamine-labeled glycopeptide fractions. One fraction (A) contains approximately eight glycose units. All of the radioactivity is at nonreducing termini and can be released by treatment with an exo-β-N-acetylglucosaminidase. A smaller N-acetyl[]glucosamine-labeled glycopeptide (B) is recovered in the elution volume expected for an asparaginyl disaccharide. Structural studies show that the labeled saccharide unit in glycopeptide B is N,N′-diacetyl[]chitobiose. The linkage between the -labeled disaccharide and the polypeptide has the properties of an N-glycosidic attachment to asparagine. Only the larger N-acetyl[]glucosamine-labeled glycopeptide (A) is found in Pronase digests of white matter membrane N-acetyl[]glucosamine-labeled glycoprotein after incubation with UDP-N-acetyl[]glucosamine. When gray matter membranes are incubated with UDP-N-acetyl[]glucosamine in the presence of tunicamycin or UMP, the labeling of glycolipid and the asparaginyl disaccharide is inhibited. UMP and tunicamycin have no effect on the transfer of N-acetyl[]glucosamine to external acceptor sites of the larger glycopeptide (A). The transfer of N,N′-diacetyl[]-chitobiose from carrier lipid to protein is observed when extensively washed membranes containing endogenous, prelabeled -labeled glycolipids are incubated in the presence or absence of unlabeled GDP-mannose. UMP treatment of the prelabeled membranes selectively discharged over 80% of the label from N-acetyl[]glucosaminyl-pyrophosphoryl-dolichol, but had no effect on the transfer of the -labeled disaccharide to protein. All of these results are concordant with transfer of N,N′-diacetylchitobiose from dolichyl diphosphate to gray matter glycoprotein. The major membrane glycoprotein labeled by the lipid-mediated []disaccharide transfer reaction has an apparent molecular weight of 24,000. Tunicamycin prevents the enzymatic labeling of the gray matter glycoprotein having an apparent molecular weight of 24,000. 相似文献
3.
(1) +/electron acceptor ratios have been determined with the oxidant pulse method for cells of denitrifying Paracoccus denitrificans oxidizing endogenous substrates during reduction of O2, NO?2 or N2O. Under optimal H+-translocation conditions, the ratios , , for reduction to N2 and for reduction to N2O were 6.0–6.3, 4.02, 5.79 and 3.37, respectively. (2) With ascorbate/N,N,N′,N′-tetramethyl-p-phenylenediamine as exogenous substrate, addition of NO?2 or N2O to an anaerobic cell suspension resulted in rapid alkalinization of the outer bulk medium. , for reduction to N2 and for reduction to N2O were ?0.84, ?2.33 and ?1.90, respectively. (3) The ratios, mentioned in item 2, were not altered in the presence of and the triphenylmethylphosphonium cation. (4) A simplified scheme of electron transport to O2, NO?2 and N2O is presented which shows a periplasmic orientation of the nitrite reductase as well as the nitrous oxide reductase. Electrons destined for NO?2, N2O or O2 pass two H+-translocating sites. The acceptor ratios predicted by this scheme are in good agreement with the experimental values. 相似文献
4.
In order to test the question if a pool of lipophilic ions may exist in black lipid membranes which cannot be detected by electrical relaxation measurements we have performed simultaneously measurements of the optical absorption of a lipophilic ion. The absorbance of membrane-bound dipicrylamine at 410 nm was measured with a sensitive spectrophotometer which can detect absorbance changes . A minimal concentration of about 6 · 1011 dipicrylamine ions per cm2 of the membrane could be detected with this instrument. The dipicrylamine concentration in the membrane obtained with the optical method is compared with the concentrations obtained from simultaneous electrical relaxation measurements. and agreed at low dipicrylamine concentrations (10?8–10?7 M in the aqueous phase) and showed saturation at higher concentrations (up to 5 · 10?6 M). In the saturation range was maximally four times higher than . The significance of this difference is discussed together with general aspects of the saturation phenomenon. 相似文献
5.
Ron L. Batstone-Cunningham Robert E. Hardy Kilian Dill 《International journal of biological macromolecules》1983,5(5):314-317
Structural studies of homozygous glycophorin AM were undertaken by monitoring the 13C methyl resonances of 13C reductively methylated glycophorin AM (contains five Lys residues, and the N-terminal Ser residues) in various forms of glycosylation. The results indicate that removal of the α-d-NeuAc residues does not affect the structure about the N-terminal Ser residue. However, removal of the fifteen O-linked oligosaccharide units results in a structural effect about the N-terminal Ser residue. Partial methylation experiments performed on native glycophorin AM and deglycosylated glycophorin AM indicate that methylation of the lysine residue(s) may influence the structure about the N-terminal Ser residue, especially in the case of deglycosylated AM. 相似文献
6.
Robert E. Hardy Marsha E. Daman Anne M. Holbrooks Kilian Dill 《International journal of biological macromolecules》1984,6(2):103-107
13C nuclear magnetic resonance (n.m.r.) spectral data for 13C reductively methylated N-terminal tryptic glycopeptides and for 13C reductively methylated N-terminal glyco-octapeptides derived from homozygous glycophorins AM and AN are presented. Their 13C chemical shift data are compared with the previously published 13C n.m.r. data for 13C reductively methylated homozygous glycophorins AM and AN in order to investigate the means of display of the MN blood determinants by these species. The pH dependence of the 13C resonances of leucine of glyco-octapeptide AN and of serine of glyco-octapepti AM indicated that only a slight structural perturbation occurs at the N-terminus when a large portion of the glycoprotein molecule is removed. However, one structural ‘state’ of 13C reductively methylated glycophorin AM is lost when the glyco-octapeptide AM is produced. The 13C resonance of leucine of glycooctapeptide AN titrated with a of 7.7 (Hill coefficient ). The 13C resonance of serine, on the other hand, exhibited an unusual pH dependence, indicating the existence of some possible steric constraints or hydrogen bonding in this molecule. In comparison to the data obtained for 13C-labelled glycooctapeptide AM molecule, the pH dependence of the chemical shift of the 13C resonance of serine of tripeptide tri-L-serine is also presented. Circular dichroism (c.d.) spectra indicated that the reductive methylation technique does not cause a large perturbation of the glycophorin A molecule. 相似文献
7.
R S Himmelwright N C Eickman E I Solomon 《Biochemical and biophysical research communications》1978,84(2):300-305
Half hemocyanin is shown to undergo a unique change at the Cu(II)?Cu(I) active site with temperature, exhibiting class II mixed valent properties at low temperature (The appearance of an intense near IR intervalence-transfer transition and a delocalized EPR spectrum). This requires a Cu(II)NNNCu(I) bridging geometry. The effects of CO coordination to half , combined with the presence of a low energy charge transfer transition, demonstrate that azide is also bridging at room temperature. Finally, half is found to be capable of coordination of a second at the copper(II) site. 相似文献
8.
Commercial [5-14C]mevalonate is shown to contain several radioactive impurities, which give artifactually high amounts of Hyamine bound, volatile acidic radioactivity when incubated with killed or living rat renal cortex slices, as compared with [5-14C]mevalonate purified either by liquid-liquid partition chromatography or through the enzymically generated by ion-exchange chromatography. The artifactual results were not diluted by incubation with increasing amounts of unlabelled mevalonate, whereas the and []cholesterol produced by rat renal cortex slices incubated with purified [5-14C]mevalonate were both diluted to the same extent by unlabelled mevalonate. It is concluded that is genuinely oxidized to , and that purification of substrate before its use is necessary. Production of and various []lipids from purified [5-14C]mevalonate, as a function of time and substrate concentration, by renal cortex and liver slices, is described. 相似文献
9.
J Remsen D Jerina H Yagi P Cerutti 《Biochemical and biophysical research communications》1977,74(3):934-940
The reaction of bacteriophage T7-DNA with the radioactive diastereomeric benzo(a)pyrene-diol-epoxides, (±) [, ]-7β,8α-dihydroxy-9α,10β-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene, and (±) [, ]-7β,8α-dihydroxy-9β,19β-epoxy-7,8,9,10-tetrahydrobenzo(1)pyrene, was investigated. Chromatographic analysis of digests of the DNA allowed the distinction of characteristic deoxynucleoside adduct peaks for the two benzo(a)pyrene-diol-epoxides. Our results, together with data from the literature, allow the identification of these adducts as mostly N2-(10-7β,8α,9α-trihydroxy-7,8,9,10-tetrahydrobenzo(a)pyreney1)deoxyguanosine and N2-(10-7β,8α,9β-trihydroxy-7,8,9,10-tetrahydrobenzo(a)pyreney1)deoxyguanosine, respectively. DNA-benzo(a)pyrene adducts with the same chromatographic properties were formed in mouse embryo fibroblasts upon treatment with benzo(a)pyrene. 相似文献
10.
J.J. Schrijen W.A.H.M. Van Groningen-Luyben H. Nauta J.J.H.H.M. De Pont S.L. Bonting 《生物化学与生物物理学报:生物膜》1983,731(2):329-337
(1) A containing membrane fraction, isolated from pig gastric mucosa, has been further purified by means of zonal electrophoresis, leading to a 20% increase in specific activity and an increase in ratio of to basal Mg2+-ATPase activity from 9 to 20. (2) The target size of , determined by radiation inactivation analysis, is 332 kDa, in excellent agreement with the earlier value of 327 kDa obtained from the subunit composition and subunit molecular weights. This shows that the Kepner-Macey factor of 6.4·1011 is valid for membrane-bound ATPases. (3) The target size of is 444 kDa, which, in connection with a subunit molecular weight of 110000, suggests a tetrameric assembly of the native enzyme. The ouabain-insensitive K+-stimulated activity has a target size of 295 kDa. (4) In the presence of added Mg2+ the target sizes of the and its phosphatase activity are decreased by about 15%, while that for the is not significantly changed. This observation is discussed in terms of a Mg2+-induced tightening of the subunits composing the molecule. 相似文献
11.
Using a rapid pH electrode, measurements were made of the flash-induced proton transport in isolated spinach chloroplasts. To calibrate the system, we assumed that in the presence of ferricyanide and in steady-state flashing light, each flash liberates from water one proton per reaction chain. We concluded that with both ferricyanide and methylviologen as acceptors two protons per electron are translocated by the electron transport chain connecting Photosystem II and I. With methyl viologen but not with ferricyanide as an acceptor, two additional protons per electron are taken up due to Photosystem I activity. One of these latter protons is translocated to the inside of the thylakoid while the other is taken up in H2O2 formation. Assuming that the proton released during water splitting remains inside the thylakoid, we compute ratios of 3 and 4 for ferricyanide and methyl viologen, respectively.In continuous light of low intensity, we obtained the same ratios. However, with higher intensities where electron transport becomes rate limited by the internal pH, the ratio approached 2 as a limit for both acceptors.A working model is presented which includes two sites of proton translocation, one between the photoacts, the other connected to Photosystem I, each of which translocates two protons per electron. Each site presents a ≈ 30 ms diffusion barrier to proton passage which can be lowered by uncouplers to 6–10 ms. 相似文献
12.
R C Warrington 《Analytical biochemistry》1974,62(1):204-216
A general procedure for the isolation of 3′-linked fragments derived from tRNA molecules is described. Purified N-2-naphthoxyacetylglycyl derivatives of the and of yeast were exhaustively digested with RNase T1 and the 3′-linked fragments (bearing the derivative) were separated from other degradation products (lacking the derivative) by stepwise chromatography on BD-cellulose. Subsequent chromatographic resolution and base-composition analysis allowed tentative identification of the 3′-terminals of and as Gp(Cp,Ap)CpCpA and Gp(Cp,Cp,Up,Ap)CpCpA, respectively. The potential utility of this procedure for development of a novel approach to nucleic acid sequence analysis is discussed. 相似文献
13.
The uptake of radiolabeled carnitine and butyrobetaine has been studied in human heart cells (CCL 27). The uptake of carnitine is 3–10-fold higher in heart cells than in fibroblasts (). The uptake of carnitine increases with temperature coefficient of 1.6 in the interval 10–20° C and with a negligible uptake at 4 and 10° C. The uptake of carnitine follows Michaelis-Menten kinetics with a of and . Carnitine uptake is suppressed 90% by NaF (24 mM). Butyrobetaine is taken up into heart cells to the same extent as carnitine with a of 5.7–17.3 μM and . Butyrobetaine inhibits competitively the uptake of carnitine and carnitine inhibits the uptake of butyrobetaine to the same extent. No conversion of radiolabeled butyrobetaine to carnitine, or carnitine to methyl choline was observed intra- or extracellulary during incubation. These data are compatible with a selective transport mechanism for carnitine which is also responsible for the uptake of butyrobetaine. 相似文献
14.
Various respiratory electron transport activities of Rhodopseudomonas capsulata were studied in membrane fragments prepared from photosynthetically grown cells of a parental strain and two terminal oxidase-defective mutant strains. The NADH and succinate oxidase activities of the mutant having a functional oxidase, M6, were considerably more sensitive to inhibition by either antimycin A or cyanide than the corresponding activities of the mutant lacking a functional oxidase, M7. The parental strain, Z-1, but not the mutants, showed biphasic inhibitory responses of NADH and succinate oxidase activities with either antimycin A or cyanide. In certain reactions no differences in inhibitor susceptibility were found among the strains tested, implying that the pathways involved were unaffected in the mutants. In this category were the actions of rotenone on NADH oxidase, antimycin A on cytochrome reductase and, in M6 and Z-1, cyanide on oxidase. These results suggest that the respiratory chain of the parental strain branches at the ubiquinone-cytochrome region into two pathways, each branch goes to a distinct terminal oxidase, and either may be blocked independently by genetic mutation. 相似文献
15.
J K Ladha P Rowell W D Stewart 《Biochemical and biophysical research communications》1978,83(2):688-696
5-hydroxylysine, an analogue of glutamate and lysine, causes production by N2-fixing A. cylindrica; it also reversibly inhibits GS activity in vitro but has no effect on alanine dehydrogenase or GOGAT. On adding 5-hydroxylysine intracellular pools of glutamine, glutamate and aspartate decrease; those of alanine and serine increase. 5-hydroxylysine alleviates the inhibitory effect of on heterocyst production and C2H2 reduction and in cultures results in heterocyst synthesis and in C2H2 reduction. The data suggest that the GS-GOGAT pathway is the sole route of importance in primary assimilation in A. cylindrica, that alone does not inhibit nitrogenase and heterocyst production, and that GS and/or a product is involved in regulating the production of both. 相似文献
16.
The isoelectric points of the blood group and gene-associated glycosyltransferases in human ovarian cyst fluids were found by isoelectric focusing to be in the pH range 9.5–10. The and transferases in serum had isoelectric points similar to those of the enzymes in cyst fluids but transferases in serum had considerably lower isoelectric points, in the pH range 6–7. The difference in the pI values of the and transferases in the serum of a donor of the genotype enabled the two enzymes to be preparatively separated by the isoelectric focusing technique. The dissimilarity in the pI values of the transferases in ovarian cyst fluids and serum samples indicates that the isoelectric point arises from a post-translational modification of the enzyme protein. 相似文献
17.
The kinetics of isotopic Na+ flows was studied in urinary bladders of toads from the Dominican Republic. Initial studies of the potential dependence of passive serosal to mucosal 22Na+ efflux demonstrated the absence of isotope interaction and/or other coupling with passive Na+ flow. The electrical current and mucosal to serosal 22Na+ influx were then measured with transmembrane potential clamped at . Subsequent elimination of active Na+ transport mucosal amiloride permitted calculation of the rates of active Na+ transport and active and passive influx and and . The results indicate that for Dominican toad bladders mounted in chambers only Na+ contributes significantly to transepithelial active ion transport; hence . was abolished at . As Δψ approached , active efflux became demonstrable. At exceeded , so that was negative. Experimental values of agreed well with theoretical values predicted by a thermodynamic formulation: . The dependence of on Δψ is curvilinear. 相似文献
18.
A potent inhibitor of activity was purified from Sigma equine muscle ATP by cation- and anion-exchange chromatography. The isolated inhibitor was identified by atomic absorption spectroscopy and proton resonance spectroscopy to be an inorganic vanadate. The isolated vanadate and a solution of V2O5 inhibit sarcolemma with an of 1 μM in the presence of 1 mM acid (EGTA), 145 mM NaCl, 6mM MgCl2, 15 mM KCl and 2 mM synthetic ATP. The potency of the isolated vanadate in increased by free Mg2+. The inhibition is half maximally reversed by 250 μM epinephrine. Equine muscle ATP was also found to contain a second inhibitor which depends on the sulfhydryl-reducing agent dithioerythritol for inhibition. This unknown inhibitor does not depend on free Mg2+ and is half maximally reversed by 2 μM epinephrine. Prolonged storage or freeze-thawing of enzyme preparations decreases the susceptibility of the to this inhibitor. The adrenergic blocking agents, propranolol and phentolamine, do not block the catecholamine reactivation. The inhibitors in equine muscle ATP also inhibit highly purified from shark rectal gland and eel electroplax. The inhibitors in equine muscle ATP have no effect on the other sarcolemmal ATPases, Mg2+-ATPase, Ca2+-ATPase and . 相似文献
19.
Short, mild treatments of sarcoplasmic reticulum vesicles with aqueous from methanol to caused an inhibition of calcium uptake and an enhancement of ATPase activity. The treatments increased both calcium-dependent (extra) ATPase activity and calcium-independent (basic) ATPase activity of vesicles. The apparent initial reaction rate of ATPase of vesicles was about twice that of control vesicles. With increasing number () of carbon atoms of the , the maximum increment of ATPase activity increased, and both the alcohol concentration () required to inhibit calcium uptake by 50% and the alcohol concentration () required to enhance ATPase activity by 50% of the maximum increment of ATPase activity decreased as follows. The ratio, to , was constant for all values. The apparent free energy of binding of the methylene groups of to sarcoplasmic reticulum vesicles was evaluated (?796 cal/mole) and compared with data from the partition of in octanol and water (?670 cal/mole). The effects of on membrane vesicles are discussed on the basis of these data. 相似文献
20.
The activity of calcium-stimulated and magnesium-dependent adenosinetriphosphatase which possesses a high affinity for free calcium (high-affinity , EC 3.6.1.3) has been detected in rat ascites hepatoma AH109A cell plasma membranes. The high-affinity had an apparent half saturation constant of for free calcium, a maximum reaction velocity of ATP hydrolyzed/mg protein per min, and a Hill number of 0.8. Maximum activity was obtained at 0.2 μM free calcium. The high-affinity was absolutely dependent on 3–10 mM magnesium and the pH optimum was within physiological range (pH 7.2–7.5). Among the nucleoside trisphosphates tested, ATP was the best substrate, with an apparent of 30 μM. The distribution pattern of this enzyme in the subcellular fractions of the ascites hepatoma cell homogenate (as shown by the linear sucrose density gradient ultracentrifugation method) was similar to that of the known plasma membrane marker enzyme alkaline phosphatase (EC 3.1.3.1), indicating that the ATPase was located in the plasma membrane. Various agents, such as K+, Na+, ouabain, KCN, dicyclohexylcarbodiimide and NaN3, had no significant effect on the activity of high-affinity . Orthovanadate inhibited this enzyme activity with an apparent half-maximal inhibition constant of 40 μM. The high-affinity was neither inhibited by trifluoperazine, a calmodulin-antagonist, nor stimulated by bovine brain calmodulin, whether the plasma membranes were prepared with or without ethylene glycol . Since the kinetic properties of the high-affinity showed a close resemblance to those of erythrocyte plasma membrane , the high-affinity of rat ascites hepatoma cell plasma membrane is proposed to be a calcium-pumping ATPase of these cells. 相似文献