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41.
42.
Ann M. G. L. De Mazière Dietrich W. Scheuermann Philip A. P. M. Aertgeerts 《The Journal of membrane biology》1987,97(2):107-115
Summary Particles and pits of freeze-fractured gap junctions are considered as complementary structures despite the frequent observations of more regular and closer spacings of pits, ascribed to plastic deformation of particle arrays. Recently, however, the noncomplementarity of pits and particles in Purkinje fibers has been reported. To ascertain the relationship between both structures, gap junctions from fixed, cryoprotected liver and myocardium were investigated using spacing and density measurements and complementary replicas.In hepatocyte gap junctions, the center-to-center distances (mean±sd) among pits, 9.57±1.49 nm, and particles, 9.70±1.77 nm, are not significantly different. Density determinations yielded a slightly higher value for the pits, (11,510±830)/m2, than for the particles, (11,230±950)/m2. In the myocardium, the spacing of the regularly arrayed pits, 9.55±1.33 nm barely exceeds the value of 9.44±1.62 nm for the particles, which show some clustering. However, the packing density for the pits, (10,090±740)/m2, appears a little higher than that of the particles (9,890±920)/m2. As density and spacing measurements provided no decisive answers, the positions of individual pits and particles of complementary junctional faces were recorded on transparent sheets and compared. In this fashion, a one-to-one correspondence between particles and pits could be established, while small discrepancies may be attributed to plastic deformation. Moreover, the collinearity of pits and particles may be suggested by the observation of a platinum grain in the center of many pits. 相似文献
43.
44.
The fluorescence of the lipophilic prbe N-phenyl-1-naphthylamine (NPN) bound to intact cells of Escherichia coli is quenched by the addition of glucose, succinate,
-lactate, pyruvate, formate and glycerol. Partial recovery of fluorescence occurs on anaerobiosis. Use of mutants with defects in the ATP synthase or the respiratory chain show that quenching of fluorescence may be energized either by ATP hydrolysis or by substrate oxidation through the respiratory chain. Permeabilization of the outer membrane by treatment of intact cells with EDTA, or use of a mutant with an outer membrane permeable to lipophilic substances, results in a more rapid binding of NPN and in a decrease in quenching observed on substrate addition. NPN binds rapidly to everted membrane vesicles, but does not respond to membrane energization. It is proposed that inner membrane energization in intact cells alters the binding or environment of NPN in the outer membrane. The fluorescence recovery which occurs on anaerobiosis has two components. One component represents a reversal of the changes which occur on membrane energization. The other component of the fluorescence change is insensitive to the uncoupler CCCP and resembles the behaviour of NPN with everted membrane vesicles. It is suggested that a portion of the fluorescence events seen with NPN involves a response of the probe to changes in the inner membrane. 相似文献
45.
Population genetics implications of the premutation hypothesis for the generation of the fragile X mental retardation gene 总被引:2,自引:0,他引:2
R. M. Winter 《Human genetics》1987,75(3):269-271
Summary The population genetics implications of the premutation hypothesis for the generation of the fragile X mental retardation gene are explored. With some broad assumptions, the consequences of the model are that (a) 50% of mothers of probands carry the premutation; (b) 6.5% of mothers of probands receive the premutation from their mothers, 18.9% from their fathers, and 24.6% as a new mutation; (c) the incidence of carriers for the full mutation equals the incidence of affected males, whereas the incidence of carriers for the premutation is 1.35 times the incidence of affected males; (d) assuming mutation rates are equal in eggs and sperm, the mutation rate from normal to premutation alleles is 1.67x10-4; (e) the expected segregation ratio in sibs of probands is 0.44, which corresponds to observed values. In addition, predictions using the premutation hypothesis of the expected segregation ratio in sibs of mothers of probands fits well with the data of Vogel and coworkers. 相似文献
46.
47.
Jürgen Markl Stefanie Winter 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1989,159(2):139-151
Summary Three murine hybridoma cell lines secreting IgG1 antibodies to 4×6 tarantula (Eurypelma californicum) hemocyanin were isolated, and the monoclonal antibodies Ec-7, Ec-8 and Ec-24 characterized by immunoblotting, immunoelectrophoresis and ELISA. WholeEurypelma hemocyanin, and the isolated subunitsa tog served as probes. For the subunits a novel, quick purification scheme on FPLC combined with immuno-affinity chromatography was established.Additionally, two cell lines secreting IgM antibodies were isolated. These antibodies showed irrelevant cross reactivities.Ec-7 strongly reacts with subunitd and weakly withb. Ec-8 and Ec-24 are specifically directed againstEurypelma subunitsa ande, respectively. The epitopes of Ec-7 and Ec-8 are sequence-dependent, whereas the Ec-24 epitope is conformation-dependent. Ec-8 and Ec-24 are specific forEurypelma hemocyanin. Ec-7 is not reactive to crustacean, centipede or gastropod hemocyanins, but binds to scorpion hemocyanin and to the immunological correlates of subunitsd andf in the hemocyanins of the spiderCupiennius salei and the xiphosuranLimulus polyphemus.In immunoblots with different polyclonal antisera,Eurypelma andAstacus hemocyanin cross-reacted with calliphorin, a larval serum protein from the blowflyCalliphora vicina. Calliphorin and chelicerate hemocyanins share the Ec-7 epitope. Sedimentation coefficients, pH stability regions, subunit size, and electron microscopical appearance of calliphorin are indiscernable from a typical 1×6 arthropod hemocyanin. This relationship is discussed in the context of hemocyanin evolution.Abbreviations
FPLC
fast performance liquid chromatography
-
PAGE
polyacrylamide gel electrophoresis
-
SDS
sodium dodecyl sulphate
A preliminary account of this work was presented in June 1987 at the annual meeting of the Deutsche Zoologische Gesellschaft at Ulm (Markl 1987a) 相似文献
48.
Mary Welch Baillargeon Raymond G. Bistline Jr. Philip E. Sonnet 《Applied microbiology and biotechnology》1989,30(1):92-96
Summary Three strains of Geotrichum candidum (ATCC 34614, NRRL Y-552 and NRRL Y-553) were examined for lipase production and activity. Variables including medium, pH, temperature, agitation rate and incubation time were examined to define the optimal culture conditions. Growth on oil in complex medium at 30°C, 300 rpm, and pH 7 produced maximal lipase activity. Fatty acid specificity of these strains and of two crude G. candidum enzyme preparations (lipase 26557 RP, Rhône Poulenc and lipase GC-4, Amano) was measured using equimolar mixtures of methyl or butyl esters of palmitic and oleic acids. The lipase from NRRL Y-553 and lipase 26557 RP displayed preferential specificity for hydrolyzing oleic acid esters, while the lipases from ATCC 34614, NRRL Y-552 and lipase GC-4 failed to discriminate between plamitic and oleic acids. 相似文献
49.
50.
Acetyl-l-carnitine as a precursor of acetylcholine 总被引:2,自引:0,他引:2
Synthesis of [3H]acetylcholine from [3H]acetyl-l-carnitine was demonstrated in vitro by coupling the enzyme systems choline acetyltransferase and carnitine acetyltransferase. Likewise, both [3H] and [14C] labeled acetylcholine were produced when [3H]acetyl-l-carnitine andd-[U-14C] glucose were incubated with synaptosomal membrane preparations from rat brain. Transfer of the acetyl moiety from acetyl-l-carnitine to acetylcholine was dependent on concentration of acetyl-l-carnitine and required the presence of coenzyme A, which is normally produced as an inhibitory product of choline acetyltransferase. These results provide further evidence for a role of mitochondrial carnitine acetyltransferase in facilitating transfer of acetyl groups across mitochondrial membranes, thus regulating the availability in the cytoplasm of acetyl-CoA, a substrate of choline acetyltransferase. They are also consistent with a possible utility of acetyl-l-carnitine in the treatment of age-related cholinergic deficits. 相似文献