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11.
The conduction of protons in different stereoisomers of dioxolane-linked gramicidin A channels
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Two different stereoisomers of the dioxolane-linked gramicidin A (gA) channels were individually synthesized (the SS and RR dimers;. Science. 244:813-817). The structural differences between these dimers arise from different chiralities within the dioxolane linker. The SS dimer mimics the helicity and the inter- and intramolecular hydrogen bonding of the monomer-monomer association of gA's. In contrast, there is a significant disruption of the helicity and hydrogen bonding pattern of the ion channel in the RR dimer. Single ion channels formed by the SS and RR dimers in planar lipid bilayers have different proton transport properties. The lipid environment in which the different dimers are reconstituted also has significant effects on single-channel proton conductance (g(H)). g(H) in the SS dimer is about 2-4 times as large as in the RR. In phospholipid bilayers with 1 M [H(+)](bulk), the current-voltage (I-V) relationship of the SS dimer is sublinear. Under identical experimental conditions, the I-V plot of the RR dimer is supralinear (S-shaped). In glycerylmonooleate bilayers with 1 M [H(+)](bulk), both the SS and RR dimers have a supralinear I-V plot. Consistent with results previously published (. Biophys. J. 73:2489-2502), the SS dimer is stable in lipid bilayers and has fast closures. In contrast, the open state of the RR channel has closed states that can last a few seconds, and the channel eventually inactivates into a closed state in either phospholipid or glycerylmonooleate bilayers. It is concluded that the water dynamics inside the pore as related to proton wire transfer is significantly different in the RR and SS dimers. Different physical mechanisms that could account for this hypothesis are discussed. The gating of the synthetic gA dimers seems to depend on the conformation of the dioxolane link between gA's. The experimental results provide an important framework for a detailed investigation at the atomic level of proton conduction in different and relatively simple ion channel structures. 相似文献
12.
Allosteric activation mechanism of the alpha1beta2gamma2 gamma-aminobutyric acid type A receptor revealed by mutation of the conserved M2 leucine
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A conserved leucine residue in the midpoint of the second transmembrane domain (M2) of the ligand-activated ion channel family has been proposed to play an important role in receptor activation. In this study, we assessed the importance of this leucine in the activation of rat alpha1beta2gamma2 GABA receptors expressed in Xenopus laevis oocytes by site-directed mutagenesis and two-electrode voltage clamp. The hydrophobic conserved M2 leucines in alpha1(L263), beta2(L259), and gamma2(L274) subunits were mutated to the hydrophilic amino acid residue serine and coexpressed in all possible combinations with their wild-type and/or mutant counterparts. The mutation in any one subunit decreased the EC(50) and created spontaneous openings that were blocked by picrotoxin and, surprisingly, by the competitive antagonist bicuculline. The magnitudes of the shifts in GABA EC(50) and picrotoxin IC(50) as well as the degree of spontaneous openings were all correlated with the number of subunits carrying the leucine mutation. Simultaneous mutation of the GABA binding site (beta2Y157S; increased the EC(50)) and the conserved M2 leucine (beta2L259S; decreased the EC(50)) produced receptors with the predicted intermediate agonist sensitivity, indicating the two mutations affect binding and gating independently. The results are discussed in light of a proposed allosteric activation mechanism. 相似文献
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Phylogenetic evidence for horizontal transmission of group I introns in the nuclear ribosomal DNA of mushroom-forming fungi 总被引:7,自引:3,他引:4
Group I introns were discovered inserted at the same position in the
nuclear small-subunit ribosomal DNA (nuc-ssu-rDNA) in several species of
homobasidiomycetes (mushroom-forming fungi). Based on conserved intron
sequences, a pair of intron-specific primers was designed for PCR
amplification and sequencing of intron-containing rDNA repeats. Using the
intron-specific primers together with flanking rDNA primers, a PCR assay
was conducted to determine presence or absence of introns in 39 species of
homobasidiomycetes. Introns were confined to the genera Panellus,
Clavicorona, and Lentinellus. Phylogenetic analyses of nuc-ssu-rDNA and
mitochondrial ssu-rDNA sequences suggest that Clavicorona and Lentinellus
are closely related, but that Panellus is not closely related to these. The
simplest explanation for the distribution of the introns is that they have
been twice independently gained via horizontal transmission, once on the
lineage leading to Panellus, and once on the lineage leading to Lentinellus
and Clavicorona. BLAST searches using the introns from Panellus and
Lentinellus as query sequences retrieved 16 other similar group I introns
of nuc-ssu-rDNA and nuclear large-subunit rDNA (nuc-lsu-rDNA) from fungal
and green algal hosts. Phylogenetic analyses of intron sequences suggest
that the mushroom introns are monophyletic, and are nested within a clade
that contains four other introns that insert at the same position as the
mushroom introns, two from different groups of fungi and two from green
algae. The distribution of host lineages and insertion sites among the
introns suggests that horizontal and vertical transmission, homing, and
transposition have been factors in intron evolution. As distinctive,
heritable features of nuclear rDNAs in certain lineages, group I introns
have promise as phylogenetic markers. Nevertheless, the possibility of
horizontal transmission and homing also suggest that their use poses
certain pitfalls.
相似文献
15.
Pim van Hooft Herbert HT Prins Wayne M Getz Anna E Jolles Sipke E van Wieren Barend J Greyling Paul D van Helden Armanda DS Bastos 《BMC evolutionary biology》2010,10(1):106
Background
The Y-chromosomal diversity in the African buffalo (Syncerus caffer) population of Kruger National Park (KNP) is characterized by rainfall-driven haplotype frequency shifts between year cohorts. Stable Y-chromosomal polymorphism is difficult to reconcile with haplotype frequency variations without assuming frequency-dependent selection or specific interactions in the population dynamics of X- and Y-chromosomal genes, since otherwise the fittest haplotype would inevitably sweep to fixation. Stable Y-chromosomal polymorphism due one of these factors only seems possible when there are Y-chromosomal distorters of an equal sex ratio, which act by negatively affecting X-gametes, or Y-chromosomal suppressors of a female-biased sex ratio. These sex-ratio (SR) genes modify (suppress) gamete transmission in their own favour at a fitness cost, allowing for stable polymorphism. 相似文献16.
Rosemary J Redfield Wendy A Findlay Janine Bossé J Simon Kroll Andrew DS Cameron John HE Nash 《BMC evolutionary biology》2006,6(1):82-15
Background
Many bacteria can take up DNA, but the evolutionary history and function of natural competence and transformation remain obscure. The sporadic distribution of competence suggests it is frequently lost and/or gained, but this has not been examined in an explicitly phylogenetic context. Additional insight may come from the sequence specificity of uptake by species such as Haemophilus influenzae, where a 9 bp uptake signal sequence (USS) repeat is both highly overrepresented in the genome and needed for efficient DNA uptake. We used the distribution of competence genes and DNA uptake specificity in H. influenzae 's family, the Pasteurellaceae, to examine the ancestry of competence. 相似文献17.
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19.
Repeated evolution of an acetate-crossfeeding polymorphism in long-term populations of Escherichia coli 总被引:1,自引:0,他引:1
Six out of 12 independent replicate populations of Escherichia coli
maintained in long-term glucose-limited continuous culture for up to
approximately 1,750 generations evolve polymorphisms maintained by acetate
crossfeeding. In all cases, the acetate-crossfeeding phenotype is
associated with semiconstitutive overexpression of acetyl CoA synthetase,
which allows for the enhanced uptake of low levels of exogenous acetate.
Mutations in the 5' regulatory region of the acetyl CoA synthetase locus
are responsible for all the acetate crossfeeding phenotypes found. These
changes were either transposable-element insertions or a single T-->A
nucleotide substitution at position -93 relative to the acs gene
translation start site.
相似文献
20.