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101.
Sequence of the precursor to rat ornithine aminotransferase deduced from a cDNA clone 总被引:19,自引:0,他引:19
The nucleotide sequence of ornithine aminotransferase mRNA from rat liver, including the entire coding and 3' untranslated regions, was determined from two overlapping cDNA clones. The mRNA encodes a precursor polypeptide of 439 amino acid residues with a molecular weight of 48,332. The deduced amino acid composition of the proposed mature enzyme sequence (residues 35 through 439) was in good agreement with that reported for the purified protein. The amino-terminal segment of the precursor corresponding to residues 1 through 34 has an overall positive charge, containing 6 basic residues and only a single acidic residue, and is postulated to be the mitochondrial leader sequence. The first 22 amino acid residues of the proposed leader sequences share 54% homology with the leader peptide of rat ornithine transcarbamylase precursor and more limited homology to the leader peptides of other nuclear-encoded mitochondrial matrix proteins. Homology was also observed between residues 286 through 362 ornithine aminotransferase precursor and a region containing the pyridoxyl phosphate binding domain of mitochondrial aspartate aminotransferase. 相似文献
102.
Howe GT; Bucciaglia PA; Hackett WP; Furnier GR; Cordonnier-Pratt MM; Gardner G 《Molecular biology and evolution》1998,15(2):160-175
The phytochrome photoreceptors play important roles in the photoperiodic
control of vegetative bud set, growth cessation, dormancy induction, and
cold-hardiness in trees. Interestingly, ecotypic differences in
photoperiodic responses are observed in many temperate- zone tree species.
Northern and southern ecotypes of black cottonwood (Populus trichocarpa
Torr. & Gray), for example, exhibit marked differences in the timing of
short-day-induced bud set and growth cessation, and these responses are
controlled by phytochrome. Therefore, as a first step toward determining
the molecular genetic basis of photoperiodic ecotypes in trees, we
characterized the phytochrome gene (PHY) family in black cottonwood. We
recovered fragments of one PHYA and two PHYB using PCR-based cloning and by
screening a genomic library. Results from Southern analyses confirmed that
black cottonwood has one PHYA locus and two PHYB loci, which we arbitrarily
designated PHYB1 and PHYB2. Phylogenetic analyses which included PHY from
black cottonwood, Arabidopsis thaliana and tomato (Solanum lycopersicum)
suggest that the PHYB/D duplications in these species occurred
independently. When Southern blots were probed with PHYC, PHYE, and PHYE
heterologous probes, the strongest bands that we detected were those of
black cottonwood PHYA and/or PHYB. These results suggest that black
cottonwood lacks members of the PHYC/F and PHYE subfamilies. Although black
cottonwood could contain additional PHY that are distantly related to known
angiosperm PHY, our results imply that the PHY family of black cottonwood
is less complex than that of other well-characterized dicot species such as
Arabidopsis and tomato. Based on Southern analyses of five black cottonwood
genotypes representing three photoperiodic ecotypes, substantial
polymorphism was detected for at least one of the PHYB loci but not for the
PHYA locus. The novel character of the PHY family in black cottonwood, as
well as the differences in polymorphism we observed between the PHYA and
PHYB subfamilies, indicates that a number of fundamental macro- and
microevolutionary questions remain to be answered about the PHY family in
dicots.
相似文献
103.
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106.
Schuster DP Brody SL Zhou Z Bernstein M Arch R Link D Mueckler M 《American journal of physiology. Lung cellular and molecular physiology》2007,292(4):L845-L851
The pathogenesis of many lung diseases involves neutrophilic inflammation. Neutrophil functions, in turn, are critically dependent on glucose uptake and glycolysis to supply the necessary energy to meet these functions. In this study, we determined the effects of p38 mitogen-activated protein kinase and hypoxia-inducible factor (HIF)-1, as well as their potential interaction, on the expression of membrane glucose transporters and on glucose uptake in murine neutrophils. Neutrophils were harvested and purified from C57BL/6 mice and stimulated with lipopolysaccharide (LPS) in the presence or absence of specific p38 and HIF-1 inhibitors. Glucose uptake was measured as the rate of [3H]deoxyglucose (DG) uptake. We identified GLUT-1 in mouse neutrophils, but neither GLUT-3 nor GLUT-4 were detected using Western blot analysis, even after LPS stimulation. LPS stimulation did not increase GLUT-1 protein levels but did cause translocation of GLUT-1 from the cell interior to the cell surface, together with a dose-dependent increase in [3H]DG uptake, indicating that glucose uptake is regulated in these cells. LPS also activated both p38 and the HIF-1 pathway. Inhibitors of p38 and HIF-1 blocked GLUT-1 translocation and [3H]DG uptake. These data suggest that LPS-induced increases in neutrophil glucose uptake are mediated by GLUT-1 translocation to the cell surface in response to sequential activation of neutrophil p38 and HIF-1alpha in neutrophils. Given that neutrophil function and glucose metabolism are closely linked, control of the latter may represent a new target to ameliorate the deleterious effects of neutrophils on the lungs. 相似文献
107.
The Glut1 glucose transporter is one of over 300 members of the major facilitator superfamily of membrane transporters. These proteins are extremely diverse in substrate specificity and differ in their transport mechanisms. The two most common features shared by many members of this superfamily are the presence of 12 predicted transmembrane segments and an amino acid motif, R-X-G-R-R, present at equivalent positions within the cytoplasmic loops joining transmembrane segments 2-3 and 8-9. The structural and functional roles of the arginine residues within these motifs in Glut1 were investigated by expression of site-directed mutant transporters in Xenopus oocytes followed by analyses of intrinsic transport activity and the membrane topology of mutant glycosylation-scanning reporter Glut1 molecules. Substitution of lysine residues for the cluster of 3 arginine residues in each of the 2 cytoplasmic pentameric motifs of Glut1 revealed no absolute requirement for arginine side chains at any of the 6 positions for transport of 2-deoxyglucose. However, removal of the 3 positive charges at either site by substitution of glycines for the arginines completely abolished transport activity as the result of a local perturbation in the membrane topology in which the cytoplasmic loop was aberrantly translocated into the exoplasm along with the two flanking transmembrane segments. Substitution of lysines for the arginines had no affect on membrane topology. We conclude that the positive charges in the R-X-G-R-R motif form critical local cytoplasmic anchor points involved in determining the membrane topology of Glut1. These data provide a simple explanation for the presence of this conserved amino acid motif in hundreds of functionally diverse membrane transporters that share a common predicted membrane topology. 相似文献
108.
Purification of galectin-3 from ovine placenta: developmentally regulated expression and immunological relevance 总被引:1,自引:1,他引:1
Iglesias MM; Rabinovich GA; Ambrosio AL; Castagna LF; Sotomayor CE; Wolfenstein-Todel C 《Glycobiology》1998,8(1):59-65
Galectins, beta-galactoside-binding lectins, are extensively distributed in
the animal kingdom and share some basic molecular properties. Galectin-3, a
member of this family, is generally associated with differentiation,
morphogenesis, and metastasis. In this study, galectin-3 was isolated from
ovine placental cotyledons round the middle of the gestation period by
lactose extraction followed by affinity chromatography on lactosyl-agarose,
and separated from galectin-1 by size exclusion chromatography on a
Superose 12 column. Under native conditions this lectin behaved as a
monomer with an apparent molecular weight of approximately 29,000 and an
isoelectric point of 9.0. The partial amino acid sequence of the peptides
obtained by tryptic digestion of this protein followed by HPLC separation
showed striking homology with other members of the galectin-3 subfamily.
Furthermore, ovine placental galectin-3 exhibited specific mitogenic
activity toward rat spleen mononuclear cells. Besides, this protein
strongly reacted with a rabbit antiserum raised against a chicken galectin.
Results obtained by Western blot analysis showed that its expression was
greatly decreased in term placenta with respect to the middle of the
gestation period, suggesting a regulated expression throughout development.
相似文献
109.
Exosomes secreted from mutant‐HIF‐1α‐modified bone‐marrow‐derived mesenchymal stem cells attenuate early steroid‐induced avascular necrosis of femoral head in rabbit 下载免费PDF全文
110.
Testing the covarion hypothesis of molecular evolution 总被引:6,自引:8,他引:6
The covarion hypothesis of molecular evolution states that the fixation of
mutations may alter the probability that any given position will fix the
next change. Tests of this hypothesis using the divergence of real
sequences are compromised because models of rate variation among sites
(e.g., the gamma version of the one-parameter equation) predict sequence
divergence values similar to those for the covarion process. This study
therefore focuses on the extent to which the varied and unvaried codons of
two well-diverged taxa are the same, because fewer are expected by the
covarion hypothesis than by the gamma model. The data for these tests are
the protein sequences of Cu, Zn superoxide dismutase (SOD) for mammals and
plants. Simulation analyses show that the covarion hypothesis makes better
predictions about the frequencies of varied and unhit positions in common
between these two taxa than does the gamma version of the one-parameter
model. Furthermore, the analysis of SOD tertiary structure demonstrates
that mammal and plant variabilities are distributed differently on the
protein. These results support the conclusions that the variable and
invariable codons of mammal and plant SODs are different and that the
covarion model explains the evolution of this protein better than the gamma
version of the one-parameter process. Unlike other models, the covarion
hypothesis accounts for rate fluctuations among positions over time, which
is an important parameter of molecular evolution.
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