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91.
Summary The equilibrium binding mechanism and kinetics of binding of diS–C3-(5) (3,3-dipropylthiodicarbocyanine iodide) to rabbit renal brush-border membrane vesicles (BBMV) were examined using steady-state and time-resolved fluorescence, and fluorescence stopped-flow methods. In aqueous solution, diS–C3-(5) exists as a monomer at concentrations <5 m with fluorescence emission peak at 670 nm (excitation 622 nm), anisotropyr=0.102, and lifetime =1.2 nsec (23°C). Upon addition of increasing BBMV (voltage clamped to 0 mV using K+/valinomycin), the 670 nm emission peak decreases, corresponding to formation of a nonfluorescent membrane dimer, and subsequently a new emission peak at 695 nm increases, corresponding to membrane monomer. Dynamic depolarization studies show that aqueous diS–C3-(5) rotation is unhindered with a rotational rateR=0.57 nsec–1 while membrane monomer is hindered with steady-state anisotropyr=0.190, lifetime =2.1 nsec,R=0.58 nsec–1 and limiting anisotropyr
=0.11. Based on equilibrium fluorescence titrations, the membrane monomer-dimer (M-D) dissociation constant,K
d=[M]2/[D][BBMV], is 0.0013, where BBMV is expressed as membrane phospholipid concentration. Three distinct kinetic processes are identified by stopped-flow experiments in which BBMV are mixed with diS–C3-(5). There is rapid binding of diS–C3-(5) to the membrane to form bound monomer with a 6-msec exponential time constant. The membrane monomer at the membrane outer surface then aggregates to form bound dimer at the outer surface with a concentration independent time constant of 30 msec. The overall dimerization reaction probably consists of a rate-limiting reorientation process (30 msec) followed by a rapid dimerization which occurs on a nanosecond time scale. Finally, there is a 0.8 to 1 sec translocation of membrane dimer between symmetric sites at the inner and outer membrane surfaces. The translocation reaction is the step which is probably sensitive to changes in transmembrane electrical potential. 相似文献
92.
Hans-B?rje Jansson A. Jeyaprakash Gerald C. Coles Nahum Marban-Mendoza Bert M. Zuckerman 《Journal of nematology》1986,18(4):570-574
Caenorhabditis elegans and Panagrellus redivivus were investigated for surface carbohydrates using fluorescent-labelled and ferritin-labelled lectins. Rhodamine-labelled Concanavalin A was specifically located in the cephalic region of both species. Rhodamine-labelled wheat germ agglutinin was located over the entire cuticle of P. redivivus but was absent on C. elegans. Rhodamine-labelled peanut agglutinin and Limax flavus agglutinin did not label nematodes of either species. Galactose and sialic acid were not detected on either species, whereas mannose-glucose residues were specifically localized in the head areas of both species. No detectable N-acetylglucosamine occurred on C. elegans, but it was evenly distributed over the cuticle surface of P. redivivus. 相似文献
93.
High-resolution proton magnetic resonance spectroscopy was used to analyze human cerebrospinal fluid obtained from patients with several neurological problems. The major metabolites measured included glucose, lactate, glutamine, citrate, inositol, acetate, creatine, creatinine, beta-hydroxybutyrate, alanine, and pyruvate. A drug vehicle, propylene glycol, was also measured. Alterations in the cerebrospinal fluid of these metabolites provided information concerning metabolism of the brain. Magnetic resonance spectroscopy offered a simple and rapid means of assessing these and other exogenous and endogenous compounds in diseases affecting the nervous system. 相似文献
94.
Human blood was sheared between rotating polyethylene disks and plasma hemoglobin measured at intervals to produce kinetic
hemolysis curves (KHC), plotted as free hemoglobin concentration vs time. The KHC produced by blood samples incubated in the
presence of penicillin, streptomycin, gentamicin, and amikacin lie always below those for control samples, indicating a reduction
in hemolysis; this reduction was greater as the drug concentration was increased. Explanations in terms of alterations in
red cell structure were sought by several characterization tests of amikacin-loaded blood samples. Drug-localization studies
demonstrated that significant fractions of the total dosage were associated with the red-cell membrane. Resistive pulse spectroscopy
was used to show how amikacin affected cell size, deformability, and osmotic fragility; results were sensitive to storage
age of the blood. In all cases, the effect of shearing was to reduce cell size, deformability, and osmotic fragility. Mechanisms
for hemolytic protection by drugs are proposed. 相似文献
95.
96.
H. J. Weigel 《Physiologia plantarum》1985,63(2):192-200
Photosynthetic CO2-fixation of mesophyll protoplasts of lambs lettuce [Valerianella locusta (L.) Betcke] was inhibited by short time exposure to Cd+. Inhibition was due to uptake of the metal ion into the protoplasts and increased with increasing Cd2+ concentrations and the time of preincubation. A 10 min pretreatment at 2 mM Cd2+ reduced CO2-fixation by 40–60%. Inhibition of photosynthesis was independent of the light intensity to which the protoplasts were exposed. Measurement of the lightinduced electrochromic pigment absorption change at 518nm and chlorophyll fluorescence studies revealed that primary photochemical reactions associated with the thylakoid membranes were not affected by the metal ion. Also, light activation of the ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) was not inhibited by Cd2+. Under rate-limiting CO2 concentrations, inhibition of CO2-fixation was smaller than at Vmax of CO2 reduction indicating that the carboxylation reaction of the Calvin cycle is not susceptible to Cd2+. Cd2+ treatment of protoplasts significantly extended the lagphase of CO2-supported O2-evolution and partly inhibited light activation of the glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13) and the ribulose-5-phosphate kinase (EC 2.7.1.19). Measurement of relative concentrations of [14C]-labeled Calvin cycle intermediates showed that Cd2+ caused a decrease in the 3-phosphoglycerate/triose phosphate ratio and an increase in the triose phosphate/ribulose-1,5-bisphosphate ratio. It is concluded that in protoplasts Cd2+ affects photosynthesis mainly at the level of dark reactions and that the site of inhibition may be localized in the regenerative phase of the Calvin cycle. 相似文献
97.
Thylakoids isolated from peas (Pisum sativum cv. Kelvedon Wonder) and phosphorylated by incubation with ATP have been compared with non-phosphorylated thylakoids in their sensitivity to photoinhibition by exposure to illumination in vitro. Assays of the kinetics of fluorescence induction at 20° C and the fluorescence emission spectra at-196° C indicate a proportionally larger decrease in fluorescence as a result of photoinhibitory treatment of non-phosphorylated compared with phosphorylated thylakoids. It is concluded that protein phosphorylation can afford partial protection to thylakoids exposed to photoinhibitory conditions.Abbreviations and symbols DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
-
F
0
Level of chlorophyll fluorescence when photosystem 2 traps are open
-
F
m
Level of chlorphyll fluorescence when photosystem 2 traps are closed
- P
Maximum level of fluorescence reached in the absence of DCMU
- PSI (II)
photosystem I(II) 相似文献
98.
Photoacoustic and fluorescence measurements of the chilling response and their relationship to carbon dioxide uptake in tomato plants 总被引:5,自引:0,他引:5
The response of tomato plants to various chilling treatments was studied using two approaches for the measurement of photosynthetic activity. One involved the use of a portable fluorometer for the measurement of in-vivo chlorophyll fluorescence, while the other employed a newly introduced photoacoustic system which allowed changes in oxygen evolution to be followed in a leaf disc. A strong correlation was found between results obtained by each system and those obtained by a conventional open gas-exchange system for the determination of CO2 uptake. Both systems of measurements could readily distinguish between the effects of chilling in the dark (at 3° C for 18 h) and chilling at high photon flux density (2000 mol m-2 s-1 for 5h at 5° C). Chilling in the dark had practically no effect on the quantum yield of oxygen evolution, chlorophyll fluorescence or CO2 uptake, while chilling at excessively high photon flux density resulted in a sharp reduction (50–70%) in the quantum yields obtained. The results support the view that photosystem II cannot be the primary site of damage by chilling in the dark, although it is significantly affected by chilling at high light intensity.Abbreviations DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- PA
photoacoustic
- PFD
photon flux density
- PSII
photosystem II 相似文献
99.
K. D. Hahn F. Buck H. Rüterjans B. K. Chernov K. G. Skryabin M. P. Kirpichnikov 《European biophysics journal : EBJ》1985,12(2):87-95
The 17 base pair operator O
R
3 oligonucleotide, which is the preferential binding site for the Cro repressor of phage , was studied by two-dimensional NMR spectroscopy. A sequential assignment procedure based on two-dimensional Nuclear Overhauser Effect (NOESY) and scalar coupling correlated (COSY) NMR spectroscopy, together with the knowledge of the oligodesoxynucleotide sequence, made it possible to assign the non-exhangeable base protons and the H1 and the H2-H2 sugar protons of the O
R
3 operator DNA. The pattern of the observed NOE connectivities is consistent with a right-handed helical DNA structure. The base and sugar proton assignments provide the necessary information for further studies of the O
R
3 operator — Cro repressor interaction.Abbreviations COSY
correlated spectroscopy
- FID
free induction decay
- NOE
nuclear Overhauser effect
- NOESY
nuclear Overhauser effect spectroscopy
- RD
relaxation delay
- TSP
sodium 3-trimethylsilyl-(2,2,3,3-2H4)propionate
- EDTA
sodium ethylendiamine tetraacetate 相似文献
100.
W. L. C. Vaz D. Hallmann R. M. Clegg A. Gambacorta M. De Rosa 《European biophysics journal : EBJ》1985,12(1):19-24
We have used the fluorescence recovery after photobleaching technique to study the translational diffusion, in L phase multibilayers of 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), of fluorescent derivatives of 1-palmitoyl-2-oleoylphosphatidylethanolamine (NBD-POPE) and a membrane-spanning phosphatidylethanolamine (NBD-MSPE). The latter derivative was prepared from a membrane-spanning glycerol-dialkyl-glycerol tetraether lipid isolated from the thermophilic and acidophilic archaebacterium Sulfolobus solfataricus. The translational diffusion was examined between about 15° and 45°C. It is shown that over this temperature range the translational diffusion coefficient for NBD-MSPE is 2/3 that for NBD-POPE which spans only one monolayer of the bilayer. The result is interpreted in terms of existing models for translational diffusion in lipid membranes.Abbreviations D
t
translational diffusion coefficient
- FRAP
fluorescence recovery after photobleaching
- MSPE
a membrane-spanning phosphatidylethanolamine derived from a glycerol-dialkyl-glycerol tetraether lipid isolated from Sulfolobus solfataricus
- NBD
4-nitrobenz-2-oxa-1,3-diazolyl
- PE
phosphatidylethanolamine
- POPC
1-palmitoyl-2-oleoylphosphatidylcholine
- POPE
1-palmitoyl-2-oleoylphosphatidylethanolamine 相似文献