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1.
The NHE-1 isoform of the Na+/H+ exchanger is excessively activated in cardiac cells during ischemia. Hence NHE-1 specific inhibitors are being developed
since they could be of beneficial influence under conditions of cardiac ischemia and reperfusion. In this study, the Cytosensor™
microphysiometer was used to measure the potency of four new drug molecules, i.e., EMD 84021, EMD 94309, EMD 96785 and HOE
642 which are inhibitors of the isoform 1 of the Na+/H+ exchanger. The experiments were performed with Chinese hamster ovary cells (CHO K1) which are enriched in the NHE-1 isoform
of the Na+/H+ antiporter. The Na+/H+ exchanger was stimulated with NaCl and the rate of extracellular acidification was quantified with the Cytosensor. The proton
exchange rate was measured as a function of the NaCl concentration in the range of 10–138 mm NaCl stimulation. The proton exchange rate followed Michaelis-Menten kinetics with a K
M
= 30 ± 4 mm for Na+. Addition of either one of the four inhibitors decreased the acidification rate. The IC50 values of the four compounds could be determined as 23 ± 7 nm for EMD 84021, 5 ± 1 nm for EMD 94309, 9 ± 2 nm for EMD 96785 and 8 ± 2 nm for HOE 642 at 138 mm NaCl, in good agreement with more elaborate biological assays. The IC50 values increased with the NaCl concentration indicating competitive binding of the inhibitor. The microphysiometer approach
is a fast and simple method to measure the activity of the Na+/H+ antiporter and allows a quantitative kinetic analysis of the proton excretion rate.
Received: 3 September 1998/Revised: 20 November 1998 相似文献
2.
The Nonrandom Location of Synonymous Codons Suggests That Reading Frame-Independent Forces Have Patterned Codon Preferences 总被引:6,自引:0,他引:6
Biased codon usage is common in eukaryotic and prokaryotic genes. Evidence from Escherichia, Saccharomyces, and Drosophila indicates that it favors translational efficiency and accuracy. However, to date no functional advantages have been identified
in the codon–anticodon interactions involving the most frequently used (preferred) codons. Here we present evidence that forces
not related to the individual codon–anticodon interaction may be involved in determining which synonymous codons are preferred
or avoided. We show that the ``off-frame' trinucleotide motif preferences inferrable from Drosophila coding regions are often in the same direction as Drosophila's ``in-frame' codon preferences, i.e., its codon usage. The off-frame preferences were inferred from the nonrandomness of
the location of confamilial synonymous codons along coding regions—a pattern often described as a context dependence of nucleotide
choice at synonymous positions or as codon-pair bias. We relied on randomizations of the location of confamilial codons that
do not alter, and cannot be influenced by, the encoded amino acid sequences, codon usage, or base composition of the genes
examined. The statistically significant congruency of in-frame and off-frame trinucleotide preferences suggests that the same
kind of reading-frame-independent force(s) may also influence synonymous codon choice. These forces may have produced biases
in codon usage that then led to the evolution of the translational advantages of these motifs as preferred codons. Under this
scenario, tRNA pool size differences between preferred and nonpreferred codons initially were evolved to track the default
overrepresentation of codons with preferred motifs. The motif preference hypothesis can explain the structuring of codon preferences
and the similarities in the codon usages of distantly related organisms.
Received: 10 November 1998 / Accepted: 23 February 1999 相似文献
3.
Nuclear and Nucleomorph SSU rDNA Phylogeny in the Cryptophyta and the Evolution of Cryptophyte Diversity 总被引:5,自引:0,他引:5
The plastid-bearing members of the Cryptophyta contain two functional eukaryotic genomes of different phylogenetic origin,
residing in the nucleus and in the nucleomorph, respectively. These widespread and diverse protists thus offer a unique opportunity
to study the coevolution of two different eukaryotic genomes within one group of organisms. In this study, the SSU rRNA genes
of both genomes were PCR-amplified with specific primers and phylogenetic analyses were performed on different data sets using
different evolutionary models. The results show that the composition of the principal clades obtained from the phylogenetic
analyses of both genes was largely congruent, but striking differences in evolutionary rates were observed. These affected
the topologies of the nuclear and nucleomorph phylogenies differently, resulting in long-branch attraction artifacts when
simple evolutionary models were applied. Deletion of long-branch taxa stabilized the internal branching order in both phylogenies
and resulted in a completely resolved topology in the nucleomorph phylogeny. A comparison of the tree topologies derived from
SSU rDNA sequences with characters previously used in cryptophyte systematics revealed that the biliprotein type was congruent,
but the type of inner periplast component incongruent, with the molecular trees. The latter is indicative of a hidden cellular
dimorphism (cells with two periplast types present in a single clonal strain) of presumably widespread occurrence throughout
cryptophyte diversity, which, in consequence, has far-reaching implications for cryptophyte systematics as it is practiced
today. 相似文献
4.
Phylogenetic Relationships of Acanthocephala Based on Analysis of 18S Ribosomal RNA Gene Sequences 总被引:3,自引:0,他引:3
García-Varela M Pérez-Ponce de León G de la Torre P Cummings MP Sarma SS Laclette JP 《Journal of molecular evolution》2000,50(6):532-540
Acanthocephala (thorny-headed worms) is a phylum of endoparasites of vertebrates and arthropods, included among the most
phylogenetically basal tripoblastic pseudocoelomates. The phylum is divided into three classes: Archiacanthocephala, Palaeacanthocephala,
and Eoacanthocephala. These classes are distinguished by morphological characters such as location of lacunar canals, persistence
of ligament sacs in females, number and type of cement glands in males, number and size of proboscis hooks, host taxonomy,
and ecology. To understand better the phylogenetic relationships within Acanthocephala, and between Acanthocephala and Rotifera,
we sequenced the nearly complete 18S rRNA genes of nine species from the three classes of Acanthocephala and four species
of Rotifera from the classes Bdelloidea and Monogononta. Phylogenetic relationships were inferred by maximum-likelihood analyses
of these new sequences and others previously determined. The analyses showed that Acanthocephala is the sister group to a
clade including Eoacanthocephala and Palaeacanthocephala. Archiacanthocephala exhibited a slower rate of evolution at the
nucleotide level, as evidenced by shorter branch lengths for the group. We found statistically significant support for the
monophyly of Rotifera, represented in our analysis by species from the clade Eurotatoria, which includes the classes Bdelloidea
and Monogononta. Eurotatoria also appears as the sister group to Acanthocephala.
Received: 12 October 1999 / Accepted: 8 February 2000 相似文献
5.
Thomas A. Gorr Barbara K. Mable Traute Kleinschmidt 《Journal of molecular evolution》1998,47(4):471-485
Phylogenetic relationships among reptiles were examined using previously published and newly determined hemoglobin sequences.
Trees reconstructed from these sequences using maximum-parsimony, neighbor-joining, and maximum-likelihood algorithms were
compared with a phylogenetic tree of Amniota, which was assembled on the basis of published morphological data. All analyses differentiated α chains into αA and αD types, which are present in all reptiles except crocodiles, where only αA chains are expressed. The occurrence of the αD chain in squamates (lizards and snakes only in this study) appears to be a general characteristic of these species. Lizards
and snakes also express two types of β chains (βI and βII), while only one type of β chain is present in birds and crocodiles.
Reconstructed hemoglobin trees for both α and β sequences did not yield the monophyletic Archosauria (i.e., crocodilians + birds) and Lepidosauria (i.e., Sphenodon+ squamates) groups defined by the morphology tree. This discrepancy, as well as some other poorly resolved nodes, might be
due to substantial heterogeneity in evolutionary rates among single hemoglobin lineages. Estimation of branch lengths based
on uncorrected amino acid substitutions and on distances corrected for multiple substitutions (PAM distances) revealed that
relative rates for squamate αA and αD chains and crocodilian β chains are at least twice as high as those of the rest of the chains considered. In contrast to
these rate inequalities between reptilian orders, little variation was found within squamates, which allowed determination
of absolute evolutionary rates for this subset of hemoglobins. Rate estimates for hemoglobins of lizards and snakes yielded
1.7 (αA) and 3.3 (β) million years/PAM when calibrated with published divergence time vs. PAM distance correlates for several speciation
events within snakes and for the squamate ↔ sphenodontid split. This suggests that hemoglobin chains of squamate reptiles
evolved ∼3.5 (αA) or ∼1.7 times (β) faster than their mammalian equivalents. These data also were used to obtain a first estimate of some
intrasquamate divergence times.
Received: 15 September 1997 / Accepted: 4 February 1998 相似文献
6.
Molecular Phylogeny of Metazoan Intermediate Filament Proteins 总被引:7,自引:0,他引:7
Andreas Erber Dieter Riemer Marc Bovenschulte Klaus Weber 《Journal of molecular evolution》1998,47(6):751-762
We have cloned cytoplasmic intermediate filament (IF) proteins from a large number of invertebrate phyla using cDNA probes,
the monoclonal antibody IFA, peptide sequence information, and various RT-PCR procedures. Novel IF protein sequences reported
here include the urochordata and nine protostomic phyla, i.e., Annelida, Brachiopoda, Chaetognatha, Echiura, Nematomorpha,
Nemertea, Platyhelminthes, Phoronida, and Sipuncula. Taken together with the wealth of data on IF proteins of vertebrates
and the results on IF proteins of Cephalochordata, Mollusca, Annelida, and Nematoda, two IF prototypes emerge. The L-type,
which includes 35 sequences from 11 protostomic phyla, shares with the nuclear lamins the long version of the coil 1b subdomain
and, in most cases, a homology segment of some 120 residues in the carboxyterminal tail domain. The S-type, which includes
all four subfamilies (types I to IV) of vertebrate IF proteins, lacks 42 residues in the coil 1b subdomain and the carboxyterminal
lamin homology segment. Since IF proteins from all three phyla of the chordates have the 42-residue deletion, this deletion
arose in a progenitor prior to the divergence of the chordates into the urochordate, cephalochordate, and vertebrate lineages,
possibly already at the origin of the deuterostomic branch. Four phyla recently placed into the protostomia on grounds of
their 18S rDNA sequences (Brachiopoda, Nemertea, Phoronida, and Platyhelminthes) show IF proteins of the L-type and fit by
sequence identity criteria into the lophotrochozoic branch of the protostomia.
Received: 2 April 1998 / Accepted: 19 June 1998 相似文献
7.
Here we describe a new short retroposon family of rodents. Like the primate Alu element consisting of two similar monomers,
it is dimeric, but the left and right monomers are different and descend from B1 and ID short retroposons, respectively. Such
elements (B1-dID) were found in the genomes of Gliridae, Sciuridae, Castoridae, Caviidae, and Hystricidae. Nucleotide sequences
of this retroposon can be assigned to several structural variants. Phylogenetic analysis of B1-dID and related sequences suggests
a possible scenario of B1-dID evolution in the context of rodent evolution.
Received: 30 August 1999 / Accepted: 20 March 2000 相似文献
8.
Aphids belonging to the three genera Tuberaphis, Glyphinaphis, and Cerataphis contain extracellular fungal symbionts that resemble endocellular yeast-like symbionts of planthoppers. Whereas the symbiont
of planthoppers has a uricase (urate oxidase; EC 1.7.3.3) and recycles uric acid that the host stores, no uric acid was found
in Tuberaphis styraci, and its fungal symbiont did not exhibit the uricase activity. However, the fungal symbionts of these aphids, including that
of T. styraci, were shown to have putative uricase genes, or pseudogenes, for the uricase. Sequence analysis of these genes revealed that
deleterious mutations occurred independently on each lineage of Glyphinaphis and Tuberaphis, while no such mutation was found in the lineage of Cerataphis. These genes were almost identical to those cloned from the symbionts of planthoppers, though the host aphids and planthoppers
are phylogenetically distant. To estimate the phylogenetic relationship in detail between the fungal symbionts of aphids and
those of planthoppers, a gene tree was constructed based on the sequences of the uricase genes including their flanking regions.
As a result, the symbionts of planthoppers and Tuberaphis aphids formed a sister group against those of Glyphinaphis and Cerataphis aphids with high bootstrap confidence levels, which strongly suggests that symbionts have been horizontally transferred from
the aphids' lineage to the planthoppers'.
Received: 29 March 2000 / Accepted: 31 May 2000 相似文献
9.
Karen Miller Clare Lynch Joanne Martin Elisabeth Herniou Michael Tristem 《Journal of molecular evolution》1999,49(3):358-366
Gypsy LTR-retrotransposons have been identified in the genomes of many organisms, but only a small number of vertebrate examples
have been reported to date. Here we show that members of this family are likely to be widespread in many vertebrate classes
with the possible exceptions of mammals and birds. Phylogenetic analyses demonstrate that although there are several distinct
lineages of vertebrate gypsy LTR-retrotransposons, the majority clusters into one monophyletic clade. Groups of fungal, plant, and insect elements were
also observed, suggesting horizontal transfer between phyla may be infrequent. However, in contrast to this, there was little
evidence to support sister relationships between elements derived from vertebrate and insect hosts. In fact, the majority
of the vertebrate elements appeared to be most closely related to a group of gypsy LTR-retrotransposons present within fungi. This implies either that at least one horizontal transmission between these two
phyla has occurred previously or that a gypsy LTR-retrotransposon lineage has been lost from insect taxa.
Received: 22 December 1998 / Accepted: 6 April 1999 相似文献
10.
Varela MF Wilson TH Rodon-Rivera V Shepherd S Dehne TA Rector AC 《The Journal of membrane biology》2000,174(3):199-205
Lactose and melibiose are actively accumulated by the wild-type Escherichia coli lactose carrier, which is an integral membrane protein energized by the proton motive force. Mutants of the E. coli lactose carrier were isolated by their ability to grow on minimal plates with succinate plus IPTG in the presence of the
toxic lactose analog β-thio-o-nitrophenylgalactoside (TONPG). TONPG-resistant mutants were streaked on melibiose MacConkey indicator plates, and red clones
were picked. These melibiose positive mutants were then streaked on lactose MacConkey plates, and white clones were picked.
Transport assays indicated that the mutants had altered sugar recognition and a defect in sugar accumulation. The mutants
had a poor apparent K
m
for both lactose and melibiose in transport. One mutant had almost no ability to take up lactose, but melibiose downhill
transport was 58% (V
max
) of normal. All of the mutants accumulated methyl-α-d-galactopyranoside (TMG) to only 8% or less of normal, and two failed to accumulate. Immunoblot analysis of the mutant lactose
carrier proteins indicated that loss of sugar transport activity was not due to loss of expression in the membrane. Nucleotide
sequencing of the lacY gene from the mutants revealed changes in the following amino acids of the lactose carrier: M23I, W151L, G257D, A295D and
G377V. Two of the mutants (G257D and G377V) are novel in that they represent the first amino acids in periplasmic loops to
be implicated with changes in sugar recognition. We conclude that the amino acids M23, W151, G257, A295 and G377 of the E. coli lactose carrier play either a direct or an indirect role in sugar recognition and accumulation.
Received: 12 October 1999/Revised: 21 December 1999 相似文献
11.
The members of the PKA regulatory subunit family (PKA-R family) were analyzed by multiple sequence alignment and clustering
based on phylogenetic tree construction. According to the phylogenetic trees generated from multiple sequence alignment of
the complete sequences, the PKA-R family was divided into four subfamilies (types I to IV). Members of each subfamily were
exclusively from animals (types I and II), fungi (type III), and alveolates (type IV). Application of the same methodology
to the cAMP-binding domains, and subsequently to the region delimited by β-strands 6 and 7 of the crystal structures of bovine
RIα and rat RIIβ (the phosphate-binding cassette; PBC), proved that this highly conserved region was enough to classify unequivocally
the members of the PKA-R family. A single signature sequence, F–G–E–[LIV]–A–L–[LIMV]–x(3)–[PV]–R–[ANQV]–A, corresponding to
the PBC was identified which is characteristic of the PKA-R family and is sufficient to distinguish it from other members
of the cyclic nucleotide-binding protein superfamily. Specific determinants for the A and B domains of each R-subunit type
were also identified. Conserved residues defining the signature motif are important for interaction with cAMP or for positioning
the residues that directly interact with cAMP. Conversely, residues that define subfamilies or domain types are not conserved
and are mostly located on the loop that connects α-helix B′ and β strand 7.
Received: 2 November 2000/Accepted: 14 June 2001 相似文献
12.
Complete chloroplast 23S rRNA and psbA genes from five peridinin-containing dinoflagellates (Heterocapsa pygmaea, Heterocapsa niei, Heterocapsa rotun-data, Amphidinium carterae, and Protoceratium reticulatum) were amplified by PCR and sequenced; partial sequences were obtained from Thoracosphaera heimii and Scrippsiella trochoidea. Comparison with chloroplast 23S rRNA and psbA genes of other organisms shows that dinoflagellate chloroplast genes are the most divergent and rapidly evolving of all.
Quartet puzzling, maximum likelihood, maximum parsimony, neighbor joining, and LogDet trees were constructed. Intersite rate
variation and invariant sites were allowed for with quartet puzzling and neighbor joining. All psbA and 23S rRNA trees showed peridinin-containing dinoflagellate chloroplasts as monophyletic. In psbA trees they are related to those of chromists and red algae. In 23S rRNA trees, dinoflagellates are always the sisters of
Sporozoa (apicomplexans); maximum likelihood analysis of Heterocapsa triquetra 16S rRNA also groups the dinoflagellate and sporozoan sequences, but the other methods were inconsistent. Thus, dinoflagellate
chloroplasts may actually be related to sporozoan plastids, but the possibility of reproducible long-branch artifacts cannot
be strongly ruled out. The results for all three genes fit the idea that dinoflagellate chloroplasts originated from red algae
by a secondary endosymbiosis, possibly the same one as for chromists and Sporozoa. The marked disagreement between 16S rRNA
trees using different phylogenetic algorithms indicates that this is a rather poor molecule for elucidating overall chloroplast
phylogeny. We discuss possible reasons why both plastid and mitochondrial genomes of alveolates (Dinozoa, Sporozoa and Ciliophora)
have ultra-rapid substitution rates and a proneness to unique genomic rearrangements.
Received: 27 December 1999 / Accepted: 24 March 2000 相似文献
13.
Saier MH 《The Journal of membrane biology》2000,175(3):165-180
Channel-forming proteins/peptides fall into over 100 currently recognized families, most of which are restricted to prokaryotes
or eukaryotes, but a few of which are ubiquitous. These proteins fall into three major currently recognized classes: (i) α-helix-type
channels present in bacterial, archaeal and eukaryotic cytoplasmic and organellar membranes, (ii) β-barrel-type porins present
in the outer membranes of Gram-negative bacterial cells, mitochondria and chloroplasts, and (iii) protein/peptide toxins targeted
to the cytoplasmic membranes of cells other than those that synthesize the toxins. High-resolution 3-dimensional structural
data are available for representative proteins/peptides of all three of these channel-forming types. Each type exhibits distinctive
features that distinguish them from the other channel protein types and from carriers. Structural, functional, and evolutionary
aspects of transmembrane channel-formers are discussed.
Received: 10 September 1999/Revised: 11 February 2000 相似文献
14.
Characterization of the Hydra Lamin and Its Gene: A Molecular Phylogeny of Metazoan Lamins 总被引:4,自引:0,他引:4
Andreas Erber Dieter Riemer Helmut Hofemeister Marc Bovenschulte Reimer Stick Georgia Panopoulou Hans Lehrach Klaus Weber 《Journal of molecular evolution》1999,49(2):260-271
We report sequences for nuclear lamins from the teleost fish Danio and six invertebrates. These include two cnidarians (Hydra and Tealia), one priapulid, two echinoderms, and the cephalochordate Branchiostoma. Combining these results with earlier data on Drosophila, Caenorhabditis elegans, and various vertebrates, the following conclusions on lamin evolution can be drawn. First, all invertebrate lamins resemble
in size the vertebrate B-type lamin. Second, all lamins described previously for amphibia, birds and mammals as well as the
first lamin of a fish, characterized here, show a cluster of 7 to 12 acidic residues in the tail domain. Since this acidic
cluster is absent from all invertebrate lamins including that of the cephalochordate Branchiostoma, it was acquired with the vertebrate lineage. The larger A-type lamin of differentiated cells must have arisen subsequently
by gene duplication and insertion of an extra exon. This extra exon of the vertebrate A-lamins is the only major change in
domain organization in metazoan lamin evolution. Third, the three introns of the Hydra and Priapulus genes correspond in position to the last three introns of vertebrate B-type lamin genes. Thus the entirely different gene
organization of the C. elegans and Drosophila Dmo genes seems to reflect evolutionary drift, which probably also accounts for the fact that C. elegans has the most diverse lamin sequence. Finally we discuss the possibility that two lamin types, a constitutively expressed
one and a developmentally regulated one, arose independently on the arthropod and vertebrate lineages.
Received: 4 February 1999 / Accepted: 1 April 1999 相似文献
15.
Cyanobacteriochrome TePixJ of Thermosynechococcus elongatus harbors phycoviolobilin as a chromophore 总被引:1,自引:0,他引:1
Cyanobacteria have several putative photoreceptors (designated cyanobacteriochromes) that are related to but distinct from the established phytochromes. The GAF domain of the phototaxis regulator, PixJ, from a thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 (TePixJ_GAF) is a cyanobacteriochrome which exhibits reversible photoconversion between a blue light-absorbing form (max = 433 nm) and a green light-absorbing form (max = 531 nm). To study the chromophore, we prepared TePixJ_GAF chromoprotein from heterologously expressed Synechocystis and performed spectral analysis after denaturation by comparing it with the cyanobacterial phytochrome Cph1 which harbors phycocyanobilin (PCB) as a chromophore. The results indicated that the chromophore of TePixJ is not PCB, but its isomer, phycoviolobilin (PVB). It is suggested that the GAF domain of TePixJ has auto-lyase and auto-isomerase activities. 相似文献
16.
Short Retroposons of the B2 Superfamily: Evolution and Application for the Study of Rodent Phylogeny
Short retroposons can be used as natural phylogenetic markers. By means of hybridization and PCR analysis, we demonstrate
that B2 retroposon copies are present only in the three rodent families: Muridae, Cricetidae, and Spalacidae. This observation
highlights the close phylogenetic relation between these families. Two novel B2-related retroposon families, named DIP and
MEN elements, are described. DIP elements are found only in the genomes of jerboas (family Dipodidae) and birch mice (family
Zapodidae), demonstrating the close relationship between these rodents. MEN element copies were isolated from the squirrel,
Menetes berdmorei, but were not detected in three other species from the family Sciuridae. The MEN element has an unusual dimeric structure:
the left and right monomers are B2- and B1-related sequences, respectively. Comparison of the B2, DIP, MEN, and 4.5S1 RNA elements revealed an 80-bp core sequence located at the beginning of the B2 superfamily retroposons. This observation
suggests that these retroposon families descended from a common progenitor. A likely candidate for this direct progenitor
could be the ID retroposon.
Received: 20 December 1996 / Accepted: 17 June 1997 相似文献
17.
This study examines the diverse maximum wavelength absorption (λmax) found in crayfishes (Decapoda: Cambaridae and Parastacidae) and the associated genetic variation in their opsin locus. We
measured the wavelength absorption in the photoreceptors of six species that inhabit environments of different light intensities
(i.e., burrows, streams, standing waters, and subterranean waters). Our results indicate that there is relatively little variation
in λmax (522–530 nm) among species from different genera and families. The existing variation did not correlate with the habitat
differences of the crayfishes studied. We simultaneously sequenced the rhodopsin gene to identify the amino acid replacements
that affect shifts in maximum wavelength absorption. We then related these to changes that correlated with shifts in λmax by reconstructing ancestral character states using a maximum-likelihood approach. Using amino acid sequences obtained from
five species (all were 301 amino acids in length), we identified a number of candidates for producing shifts of 4 to 8 nm
in λmax. These amino acid replacements occurred in similar regions to those involved in spectral shifts in vertebrates.
Received: 12 March 1997 / Accepted: 3 June 1997 相似文献
18.
Calcium-induced fusion of liposomes was studied with a view to understand the role of membrane tension in this process. Lipid
mixing due to fusion was monitored by following fluorescence of rhodamine-phosphatidyl-ethanolamine incorporated into liposomal
membrane at a self-quenching concentration. The extent of lipid mixing was found to depend on the rate of calcium addition:
at slow rates it was significantly lower than when calcium was injected instantly. The vesicle inner volume was then made
accessible to external calcium by adding calcium ionophore A23187. No effect on fusion was observed at high rates of calcium
addition while at slow rates lipid mixing was eliminated. Fusion of labeled vesicles with a planar phospholipid membrane (BLM)
was studied using fluorescence microscopy. Above a threshold concentration specific for each ion, Ca2+, Mg2+, Cd2+ and La3+ induce fusion of both charged and neutral membranes. The threshold calcium concentration required for fusion was found to
be dependent on the vesicle charge, but not on the BLM charge. Pretreatment of vesicles with ionophore and calcium inhibited
vesicle fusion with BLM. This effect was reversible: chelation of calcium prior to the application of vesicle to BLM completely
restored their ability to fuse. These results support the hypothesis that tension in the outer monolayer of lipid vesicle
is a primary reason for membrane destabilization promoting membrane fusion. How this may be a common mechanism for both purely
lipidic and protein-mediated membrane fusion is discussed.
Received: 27 September 1999/Revised: 22 March 2000 相似文献
19.
20.
A Novel Family of Ubiquitous Heavy Metal Ion Transport Proteins 总被引:33,自引:0,他引:33
We describe a novel diverse family of metal ion transporter (CDF) proteins (the cation diffusion facilitator (CDF) family)
with members occurring in both prokaryotes and eukaryotes. Thirteen sequenced protein members of the CDF family have been
identified, several of which have been shown to transport cobalt, cadmium and/or zinc. All members of the CDF family possess
six putative transmembrane spanners with strongest conservation in the four N-terminal spanners, and on the basis of the analyses,
we present a unified structural model. Members of the family are shown to exhibit an unusual degree of size variation, sequence
divergence, and differences in cell localization and polarity. The phylogenetic tree for the CDF family reveals that prokaryotic
and eukaryotic proteins cluster separately. It allows functional predictions for some uncharacterized members of this family.
A signature sequence specific for the CDF family is derived.
Received: 15 July 1996/Revised: 21 October 1996 相似文献