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1.
2.
Trimethyloxonium modification of single batrachotoxin-activated sodium channels in planar bilayers. Changes in unit conductance and in block by saxitoxin and calcium 总被引:13,自引:9,他引:4
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Single batrachotoxin-activated sodium channels from rat brain were modified by trimethyloxonium (TMO) after incorporation in planar lipid bilayers. TMO modification eliminated saxitoxin (STX) sensitivity, reduced the single channel conductance by 37%, and reduced calcium block of inward sodium currents. These effects always occurred concomitantly, in an all-or-none fashion. Calcium and STX protected sodium channels from TMO modification with potencies similar to their affinities for block. Calcium inhibited STX binding to rat brain membrane vesicles and relieved toxin block of channels in bilayers, apparently by competing with STX for the toxin binding site. These results suggest that toxins, permeant cations, and blocking cations can interact with a common site on the sodium channel near the extracellular surface. It is likely that permeant cations transiently bind to this superficial site, as the first of several steps in passing inward through the channel. 相似文献
3.
Zusammenfassung Kanarienvogel (Serinus) und Stieglitz (Carduelis) zeigen für das Enzym Glucose-6-Phosphat-Dehydrogenase in der Stärkegelektrophorese jeweils nur eine Bande, die aber verschiedene Positionen einnehmen. Interspecieshybriden beider Geschlechter wurden aus der natürlichen Kreuzung Kanarienvogel (f)xStieglitz (m) erhalten; die Identifizierung des Geschlechts der äußerlich ähnlichen Hybriden erfolgte durch Chromosomenanalyse. Die Hybriden zeigen in beiden Geschlechtern ein identisches Elektrophoresemuster, das die gleiche Zone umfaßt wie bei dem in vitro-Gemisch aus Gewebehomogenaten der Ausgangsarten. Entsprechende Befunde liefert die Analyse der 6-Phosphogluconatdehydrogenase. Hieraus ist zu schließen, daß beide Enzyme von autosomalen Genen determiniert werden.
Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.
Wesentliche Teile dieser Arbeit werden von Herrn G. Oser als Dissertation der Medizinischen Fakultät der Universität Freiburg i. Br. vorgelegt. 相似文献
Autosomal determination of glucose-6-phosphate dehydrogenase in birdsStudies on interspecific hybrids of serinus and carduelis (family: Fringillidae)
Summary The electrophoretic mobility of the single G-6-PD-band in Serinus and Carduelis is different. The natural cross Serinus (f)xCarduelis (m) results in viable hybrids of both sexes, as confirmed by chromosome analysis. The isoenzymes of G-6-PD are indentical in both sexes of the hybrids, exhibiting a broad diffuse pattern comprising the same zone as obtained from an in vitro mixture of parental homogenates. Related findings are reported on the enzyme 6-PGD. It is concluded that both enzymes are autosomally determined in these species.
Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.
Wesentliche Teile dieser Arbeit werden von Herrn G. Oser als Dissertation der Medizinischen Fakultät der Universität Freiburg i. Br. vorgelegt. 相似文献
4.
Summary The conditions for optimum incorporation of radioactive amino acids into proteins of cultured postimplantation mouse embryos
were investigated under the aspect of using these proteins for two-dimensional electrophoretic separations and fluorography.
The aim was to obtain highly radioactively labeled proteins under conditions as physiological as possible. Mouse embryos of
Days 8, 10, and 11 of gestation were cultured in Tyrode’s solution. Incubation time and concentration of [3H (or14C)]amino acids in the culture medium were varied over a broad range. Embryos were prepared with placenta and yolk sac or without
any embryonic envelopes. After culturing, the physiologic-morphologic state of the embryos was registered on the basis of
several criteria. The radioactivity taken up by the total protein of each embryo was determined and calculated in disintegrations
per minute per milligram protein per embryo. To approach our aim, embryos of different developmental stages had to be cultured
under different conditions. A good compromise for Day-8, Day-10, and Day-11 embryos was: embryos prepared with yolk sac (opened)
and placenta, 150 μCi radioactive amino acids added per milliliter medium, incubation for 4 to 5 h. For maximum labeling of
proteins it is advisable to culture Day-10 embryos without embryonic envelopes under particular conditions.
This work was supported by grants from the Deutsche Forschungsgemeinschaft awarded to the project K1 237/3-2 (Systematic analysis
of cell proteins). 相似文献
5.
Restoration of membrane incorporation of an Escherichia coli outer membrane protein (OmpA) defective in membrane insertion 总被引:2,自引:0,他引:2
M Klose F J?hnig I Hindennach U Henning 《The Journal of biological chemistry》1989,264(36):21842-21847
The mechanism of sorting, to the outer membrane, of the 325-residue Escherichia coli protein OmpA has been investigated. It is thought to traverse the membrane eight times in antiparallel beta-strands, forming an amphiphilic beta-barrel which encompasses residues 1 to about 170; the COOH-terminal moiety is periplasmic. A mutant, carrying the substitutions Leu164----Pro and Val166----Asp within the last beta-strand (residues 160-170), has been described which was unable to assemble in the membrane (Klose, M., MacIntyre, S., Schwarz, H., and Henning, U. (1988) J. Biol. Chem. 263, 13297-13302). Linkers were inserted between the codons for residues 164 and 165 of the mutant protein. Of 13 different genes recovered, five encoded proteins which had regained the ability to assemble in the membrane. The properties of the mutant proteins, together with a structure prediction method, indicate the following rules for the final beta-strand to be compatible with, or possibly initiate, membrane insertion: (i) it must be amphiphilic or hydrophobic while its primary structure as such is fairly unimportant, (ii) it must extend over at least 9 residues, and (iii) it must not contain a proline residue around its center. One of the genes recovered coded for OmpA up to residue 164 and then followed by 10 linker-encoded residues. This 174-residue polypeptide was assembled in the membrane but did not, in contrast to all other proteins, expose sites sensitive to trypsin at the inner face of the membrane. This behavior agrees perfectly well with the OmpA model. 相似文献
6.
Relative apparent synapomorphy analysis (RASA). I: The statistical measurement of phylogenetic signal 总被引:10,自引:9,他引:1
We have developed a new approach to the measurement of phylogenetic signal
in character state matrices called relative apparent synapomorphy analysis
(RASA). RASA provides a deterministic, statistical measure of natural
cladistic hierarchy (phylogenetic signal) in character state matrices. The
method works by determining whether a measure of the rate of increase of
cladistic similarity among pairs of taxa as a function of phenetic
similarity is greater than a null equiprobable rate of increase. Our
investigation of the utility and limitations of RASA using simulated and
bacteriophage T7 data sets indicates that the method has numerous
advantages over existing measures of signal. A first advantage is
computational efficiency. A second advantage is that RASA employs known
methods of statistical inference, providing measurable sensitivity and
power. The performance of RASA is examined under various conditions of
branching evolution as the number of characters, character states per
character, and mutations per branch length are varied. RASA appears to
provide an unbiased and reliable measure of phylogenetic signal, and the
general approach promises to be useful in the development of new techniques
that should increase the rigor and reliability of phylogenetic estimates.
相似文献
7.
The dynamic properties of water in phosphatidylcholine lipid/water dispersions have been studied, applying a combination of 2H-NMR techniques (quadrupole splitting and spin-lattice relaxation time) and self-diffusion measurements using pulsed field gradient (PFG) 1H-NMR. The hydration properties of POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine) were compared with those of DOPC (1,2-dioleoyl-sn-glycero-3-phosphatidylcholine) and EYL (egg yolk phosphatidylcholine (lecithin)). A model is presented that assumes an exponentially decaying influence of the bilayer surface on water dynamics as well as on water orientation with increasing hydration. This assumption is based on an exponentially decaying hydration potential which results from direct lipid-water and water-water interactions. The model describes successfully the experimental data for a large water concentration range, especially at low hydration, where other models failed. With the exception of a small fraction of water which is significantly influenced by the surface in slowing down the mobility, the interbilayer water has isotropic, free water characteristics in terms of correlation times and molecular order. Hydration properties of POPC are comparable with those of EYL but differ from DOPC. At very low water content the correlation times of headgroup segmental reorientation and water are similar, indicating a strong coupling of this water to the lipid lattice. The hydration properties of the three lipids studied are explained in terms of slightly different headgroup conformations due to different lateral packing of the molecules by their fatty-acid chain composition. 相似文献
8.
9.
Variation in heat-shock proteins among species of desert fishes (Poeciliidae, Poeciliopsis) 总被引:4,自引:0,他引:4
Analysis of the heat-shock proteins (hsps) of six closely related species
of Poeciliopsis demonstrated the existence of biochemical diversity in the
hsp100, hsp70, hsp60, and hsp30 protein families among species. Each
species expressed five to seven hsp70-related isoforms. Constitutive 70-kD
isoforms were identical among species, but four different patterns of
heat-inducible isoforms were seen in these six species. Members of the
hsp70 family of molecular chaperones are included among the most highly
conserved proteins known, and the possibility of variation in hsp70 among
closely related species has rarely been addressed. The hsp30 family is
known to be less conserved than the hsp70 family, and, as expected, the
Poeciliopsis hsp30 patterns showed more variation. Most of the hsp30
isoforms characteristic of a particular species were unique to that
species. Hsp100 and hsp60 were identical in five of the species, but
alternate isoforms were found in P. monacha. The small size and limited
geographical distribution of the P. monacha population have probably
contributed to the uniqueness of the monacha pattern. Two of the species
were shown to acquire thermotolerance, the ability to withstand normally
lethal temperatures when subjected to a gradual temperature increase.
Rapid-heating protocols commonly used to establish critical thermal maxima
of organisms do not include this inducible component of thermoresistance
and therefore do not adequately assess an organism's capacity to withstand
thermal stress.
相似文献
10.
The translocation of secretory proteins derived from a Gram-positive (Staphylococcus hyicus prolipase) or a Gram-negative (Escherichia coli pre-OmpA protein) bacterium across the cytoplasmic membrane was studied in E. coli and Bacillus subtilis. in both microorganisms, the prolipase was found to be secreted across the plasma membrane when either the pre-prolipase signal peptide (38 amino acids in length) or the pre-OmpA signal peptide (21 amino acids in length) was used. Expression of the gene encoding the authentic pre-OmpA protein in B. subtilis resulted in the translocation of mature OmpA protein across the plasma membrane. Processing of the OmpA precursor in B. subtilis required the electrochemical potential and was sensitive to sodium azide, suggesting that the B. subtilis SecA homologue was involved in the translocation process. The mature OmpA protein, which was most likely present in an aggregated state, was fully accessible to proteases in protoplasted cells. Therefore, our results clearly demonstrate that an outer membrane protein can be secreted by B. subtilis, supporting the notion that the basic mechanism of protein translocation is highly conserved in Gram-positive and Gram-negative bacteria. 相似文献