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131.
Shiori Tamamizu A. Paul Todd Mark G. McNamee 《Cellular and molecular neurobiology》1995,15(4):427-438
Summary 1. Site directed mutagenesis was used to alter the structure ofTorpedo californica nicotinic acetylcholine receptor (nAChR) and to identify amino acid residues which contribute to noncompetitive inhibition by quinacrine. Mutant receptors were expressed inXenopus laevis oocytes injected within vitro synthesized mRNA and the whole cell currents induced by acetylcholine (ACh) were recorded by two electrode voltage clamp.2. A series of mutations of a highly conserved Arg at position 209 of the subunit ofTorpedo californica nAChR revealed that positively charged amino acids are required for functional receptor expression. Mutation of Arg to Lys (R209K) or His (R209H) at position 209 shifted the EC50 for ACh slightly from 5µM to 12µM and increased the normalized maximal channel activity 8.5-and 3.2-fold, respectively.3. These mutations altered the sensitivity of nAChR to noncompetitive inhibition by quinacrine. The extent of inhibition of ion channel function by quinacrine was decreased as pH increased in both wild type and mutant nAChR suggesting that the doubly charged form of quinacrine was responsible for the inhibition.4. Further mutations at different positions of the subunit suggest the contribution of Pro and Tyr residues at positions 211 and 213 to quinacrine inhibition whereas mutationsI210A andL212A did not have any effects. None of these mutations changed the sensitivity of nAChR to inhibition by a different noncompetitive inhibitor, chlorpromazine.5. These findings support a hypothesis that the quinacrine binding site is located in the lumen of the ion channel. In addition, the quantitative effect of point mutations at alternate positions on the sensitivity of quinacrine inhibition suggests that the secondary structure at the beginning of M1 region might be sheet structure. 相似文献
132.
Direct sequence analysis of proteins by in-source fragmentation during delayed ion extraction. 总被引:1,自引:1,他引:0
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J. J. Lennon K. A. Walsh 《Protein science : a publication of the Protein Society》1997,6(11):2446-2453
Continuous segments of amino acid sequence information as long as 41 residues have been deduced by interpretation of matrix-assisted laser desorption/ionization-generated ion signals dominated by Cn fragmentation within the ion source of a linear time-of-flight mass spectrometer utilizing delayed ion extraction. The technique has been applied successively to five proteins of mass 12.2 kDa to 18.3 kDa, yielding segments of continuous sequence as long as 41 residues without the need for prior proteolytic fragmentation. Intact crosslinks such as disulfides or heme linkages interrupt the generation of these data. 相似文献
133.
Physical effects of negative air ions in a wet sauna 总被引:10,自引:0,他引:10
I. Watanabe Hiroshi Noro Yoshinori Ohtsuka Yukio Mano Yuko Agishi 《International journal of biometeorology》1997,40(2):107-112
The physical effects of negative air ions on humans were determined in an experimental sauna room equipped with an ionizer.
Thirteen healthy persons took a wet sauna bath (dry bulb temperature 42° C, relative humidity 100%, 10 min exposure) with
or without negative air ions. The subjects were not told when they were being exposed to negative air ions. There were no
differences in the moods of these persons or changes in their blood pressures between the two saunas. The surface temperatures
of the foreheads, hands, and legs in the sauna with negative ions were significantly higher than those in the sauna without
ions. The pulse rates and sweat produced in the sauna with ions were singificantly higher than those in the sauna without
ions. The results suggest that negative ions may amplify the effects on humans of the sauna.
Received: 31 March 1995 / Revised: 25 July 1995 / Accepted: 26 July 1996 相似文献
134.
Previous studies in our laboratory have shown that Na absorption across the porcine endometrium is stimulated by PGF2α and cAMP-dependent activation of a barium-sensitive K channel located in the basolateral membrane of surface epithelial cells.
In this study, we identify and characterize this basolateral, barium-sensitive K conductance. Porcine uterine tissues were
mounted in Ussing chambers and bathed with KMeSO4 Ringer solution. Amphotericin B (70 μm) was added to the luminal solution to permeabilize the apical membrane and determine the current-voltage relationship of
the basolateral K conductance after activation by 100 μm CPT-cAMP. An inwardly rectifying current was identified which possessed a reversal potential of −53 mV when standard Ringer
solution was used to bathe the serosal surface. The K:Na selectivity ratio was calculated to be 12:1. Administration of 5
mm barium to the serosal solution completely inhibited the current activated by cAMP under these conditions. In addition to
these experiments, amphotericin-perforated whole cell patch clamp recordings were obtained from primary cultures of porcine
surface endometrial cells. The isolated cells displayed an inwardly rectifying current under basal conditions. This current
was significantly stimulated by CPT-cAMP and blocked by barium. These results together with our previous studies demonstrate
that cAMP increases Na absorption in porcine endometrial epithelial cells by activating an inwardly rectifying K channel present
in the basolateral membrane. Similar patch clamp experiments were conducted using cells from a human endometrial epithelial
cell line, RL95-2. An inwardly rectifying current was also identified in these cells which possessed a reversal potential
of −56 mV when the cells were bathed in standard Ringer solution. This current was blocked by barium as well as cesium. However,
the current from the human cells did not appear to be activated by cAMP, indicating that distinct subtypes of inwardly rectifying
K channels are present in endometrial epithelial cells from different species.
Received: 6 February 1997/Revised: 10 July 1997 相似文献
135.
YANG Jianbo WU Lijun LI Li WU JiadaoYU Zengliang XU Zhihong 《中国科学:生命科学英文版》1997,40(1):107-112
While M13mp18 double-stranded DNA was irradiated with ion beam, and transfected intoE. coli JM103, a decrease of transfecting activity was discovered. The lacZ- mutation frequency at 20% survival could reach (3.6–16.8) × 104, about 2, 3–10 times that of unirradiated M13DNA. Altogether, 27 IacZ-mutants were selected, 10 of which were used for sequencing. 7 of the sequenced mutants show base changes in 250-bp region
examined (the remaining 3 mutants probably have base changes outside the regions sequenced). 5 of the base-changed mutants
contain more than one mutational base sites (some of them even have 5–6 mutational base sites in 250-bp region examined);
this dense distribution of base changes in polysites has seldom been seen in X-rays, Y-rays or UV induced DNA mutations. Our
experiments also showed that the types of base changes include transitions(50%), transversions (45%) and deletion (5%); no
addition or duplication was observed. The transitions were mainly C→T and A→G; the transversions were mainly C→A and C→G.
The mutations involving cytosine residue (in the template strand) constitute about 60% of all the base changes observed. In
comparison with the surrounding sequences of mutational base sites, the base located between TG and CT is found to be easily
substituted. 相似文献
136.
Ilkka Havukkala Hideaki Ichimura Yoshiaki Nagamura Takuji Sasaki 《Plant Molecular Biology Reporter》1995,13(1):38-55
A total of 687 DNA sequence accessions from the Mendel database (release 1.04, 3 November 1994) assigned standardized designations
for plant genes and gene products were used in aBLAST similarity search of 7557 rice partial cDNA sequences and 287 other rice sequences from the Japanese Rice Genome Research
Program. We describe procedures for data manipulation, import and export from and to Macintosh and Unix, and the use of 4th
Dimension relational database management system (RDBMS) in data processing. Altogether 275 sequences showed strong similarity
hits. Using the CPGN nomenclature, we assign putative designations for genes and gene products. Assignments include representatives
of 26 gene products, including 58 cDNA sequences similar to α-tubulins (TubA), 23 similar to β-tubulins (TubB) and 51 similar to cytosolic subunit C of glyceraldehyde-3-phosphate dehydrogenase (NAD) (GapC). The results of the similarity searches are listed and are also available electronically. The assignments have been submitted
to the CPGN working groups for verification and for later inclusion in the GenBank/EMBL/DDBJ sequence databases, which will
include the standardized designations in the accession data fields.
Member of the ISPMB Commission on Plant Gene Nomenclature, representing the Rice Genome Research Program of Japan.
Reprint requests to T. Sasaki. 相似文献
137.
Secondary metabolic-energy-generating systems generate a proton motive force (pmf) or a sodium ion motive force (smf) by a
process that involves the action of secondary transporters. The (electro)chemical gradient of the solute(s) is converted into
the electrochemical gradient of protons or sodium ions. The most straightforward systems are the excretion systems by which
a metabolic end product is excreted out of the cell in symport with protons or sodium ions (energy recycling). Similarly,
solutes that were accumulated and stored in the cell under conditions of abundant energy supply may be excreted again in symport
with protons when conditions become worse (energy storage). In fermentative bacteria, a proton motive force is generated by
fermentation of weak acids, such as malate and citrate. The two components of the pmf, the membrane potential and the pH gradient,
are generated in separate steps. The weak acid is taken up by a secondary transporter either in exchange with a fermentation
product (precursor/product exchange) or by a uniporter mechanism. In both cases, net negative charge is translocated into
the cell, thereby generating a membrane potential. Decarboxylation reactions in the metabolic breakdown of the weak acid consume
cytoplasmic protons, thereby generating a pH gradient across the membrane. In this review, several examples of these different
types of secondary metabolic energy generation will be discussed. 相似文献
138.
Two series of amino-modified silicate gels prepared by sol-gel processing were used to absorb Cu(II), Ni(II), Co(II), Mn(II) and Cr(III) from aqueous solutions. These easily prepared sorbents with various content of primary amino groups in series (A) or primary and secondary amino groups in series (AA) have reasonable stability. The gel composition, time and concentration dependence of the uptake of the metal ions by these materials were studied systematically. These materials would be further used as supports to disperse catalytically active phases by conventional wet chemical procedures. Apart from this they demonstrate potential for the preconcentration aid for transition metal analysis. 相似文献
139.
140.
Calcification in aquatic plants 总被引:1,自引:0,他引:1
MICHAEL A. BOROWITZKA 《Plant, cell & environment》1984,7(6):457-466
Abstract. The CaCO3 deposits of aquatic plants may be intra-, inter- and extracellular. Calcification is mainly the result of photosynthetic CO2 or HCO− 3 assimilation. This raises the local pH and CO2− 3 concentration resulting from shifts in the dissolved inorganic carbon equilibrium, due to either net CO2 depletion as in Halimeda or localized OH− efflux (or H+ influx) as in Chara. The plant cell wall may be important in CaCO3 nucleation by acting as an epitaxial substratum or template, or by creating a microenvironment enriched in Ca2+ compared to Mg2+ . Hypotheses on the reason for the lack of calcification in many aquatic plants are presented. 相似文献