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971.
Geyer J Godoy JR Petzinger E 《Biochemical and biophysical research communications》2004,316(2):300-306
In this study, a novel sodium-dependent organic anion transporter (Soat) was identified. Soat is expressed in rat brain, heart, kidney, lung, muscle, spleen, testis, adrenal gland, small intestine, and colon. The Soat protein consists of 370 amino acids and shows 42% and 31% overall amino acid sequence identity to the ileal sodium-dependent bile acid transporter (Isbt) and the Na(+)/taurocholate cotransporting polypeptide (Ntcp), respectively. Soat is predicted to have nine transmembrane domains, with an N-terminus outside the cell and an intracellular C-terminus. The Soat gene is localized on chromosome 14 and is coded by six exons mapped in region 14p22. When expressed in Xenopus laevis oocytes, Soat shows transport function for estrone-3-sulfate (Km = 31 microM, Vmax = 5557 fmol/oocyte/30 min) and dehydroepiandrosterone sulfate (Km = 30 microM, Vmax = 5682 fmol/oocyte/30 min). Soat does not transport taurocholate, estradiol-17beta-glucuronide, nor ouabain. 相似文献
972.
Mitochondrial dysfunction in mouse oocytes results in preimplantation embryo arrest in vitro 总被引:3,自引:0,他引:3
Oocyte mitochondrial dysfunction has been proposed as a cause of high levels of developmental retardation and arrest that occur in human preimplantation embryos generated using assisted reproductive technology in the treatment of some causes of female infertility. To investigate this, a model of mitochondrial dysfunction was developed in mouse oocytes using a method of photosensitization of the mitochondrion-specific dye, rhodamine-123. After in vitro fertilization, dye-loaded and photosensitized oocytes showed developmental arrest in proportion to irradiation time. Morphological and metabolic assessments of zygotes indicated an increase in mitochondrial permeability that subsequently resulted in apoptotic degeneration. Development was partially restored by inhibition of mitochondrial permeability transition pore formation by oocyte pretreatment with cyclosporin A. Oocyte mitochondria are therefore physiological regulators of early embryo development and potential sites of pathological insult that may perturb oocyte and subsequent preimplantation embryo viability. These findings have important implications for the treatment of clinically infertile women using assisted reproductive technologies. 相似文献
973.
Böhm E Voglauer R Steinfellner W Kunert R Borth N Katinger H 《Biotechnology and bioengineering》2004,88(6):699-706
One of the major problems in the biotechnology industry is the selection of cell lines well suited for production of biopharmaceutical proteins. Usually, the most important selection criterion is the cell specific production rate. Nevertheless, a good producer cell line should have a number of additional advantageous properties, which allow the cell line to perform well in the type of bioreactor chosen for the process. However, the time and work required to select for high production rates as well as the lack of methods to specifically select for other cellular properties, usually prevents researchers from including such criteria into their screening program.With the Single Cell Secretion Assay it is possible to measure the specific production rates of individual cells by catching secreted product in an artificial matrix applied to the cell surface. Flow cytometric cell sorting then allows selection of rare cells with high production rates, which occur at frequencies as low as 10(-6). By combining this method with culture conditions that bring out a desired cellular property, we were able to isolate subclones with similar production rates, but improved performance from a recombinant Chinese hamster ovary cell line producing a human monoclonal antibody. The two desired cellular properties screened for were a non-growth associated production kinetic and improved stability in the absence of selective pressure. 相似文献
974.
Epoxidation of C-branched glycals: unexpected stereochemical results and their theoretical rationale
This paper describes the synthesis of C-3 methyl-branched glycosides by epoxidation of partially unblocked L-configured glycals. The stereochemical result depends on the orientation of the allylic hydroxyl group. A theoretical explanation is presented, based on the conformational preferences of the respective glycal half-chair conformations that were estimated by applying the BP density functional and a valence triple-zeta basis set. 相似文献
975.
Environmental influences on signal transduction through membranes: a retrospective mini-review 总被引:1,自引:0,他引:1
Helmreich EJ 《Biophysical chemistry》2003,100(1-3):519-534
This mini-review is addressed to the question how the membranous environment may affect traffic of receptors and signalling from membrane-anchored receptors on the outside of cells to transducers and targets on the inside. Signal transduction by membrane-anchored receptors to the interior of the cell and eventually to the genome is a central issue in cellular regulation. In this context the role of membrane fluidity and of the cytoskeleton in restricting the mobility of proteins are discussed and the evidence for a structural order in membranes which could limit the mobility of proteins is scrutinised. 相似文献
976.
Mutational analysis of the human FATE gene in 144 infertile men 总被引:2,自引:0,他引:2
Olesen C Silber J Eiberg H Ernst E Petersen K Lindenberg S Tommerup N 《Human genetics》2003,113(3):195-201
The FATE gene maps to Xq28 where one case of a translocation breakpoint has been found in an infertile man. Moreover, the FATE promoter contains a putative SF-1-binding site, and FATE has been proposed as representing a target gene of SF-1 in testicular development or germ cell differentiation. This study presents a complete mutational screening of the FATE gene in a random group of 144 infertile males. Four polymorphisms and two mutations were found. Three of the polymorphisms, viz., 741CT, 905AC, and 3985CT, occurred in exon 5 and intron 2 and did not alter the deduced polypeptide. One polymorphism resulted in the conservative amino acid exchange, A10 V, in 16.0% of the patients. This substitution occurred with similar frequencies in the control groups, indicating that the mutation does not affect fertility in men or women. The two mutations caused the non-conservative amino acid substitutions S125R (patient 1) and I34T (patient 2). A family study (patient 1) revealed, however, that S125R was inherited and that a fertile male family member carried the mutation. Patient 2 did not have relevant family members who could be examined. Thus, this study has shown that only 1.4% of infertile men have mutations in the FATE gene, and that some of these mutations do not singly cause infertility. Hence, FATE may not play an important role in the disease-state of infertile men attending fertility clinics. However, FATE mutations cannot be excluded as being a contributing factor in some cases of male infertility. 相似文献
977.
Huber K Roesler U Muscher A Hansen K Widiyono I Pfeffer E Breves G 《American journal of physiology. Regulatory, integrative and comparative physiology》2003,284(2):R413-R421
The rapid development of precocial goats in the first weeks after birth requires an adequate adaptation of phosphate transport systems to maintain the P homeostasis at each developmental stage. Here we examined the age-related development of Na+-Pi transport systems in small intestines, kidneys, and parotid glands of goats. Kinetic parameters were determined by brush-border membrane vesicle uptake studies, and relative expression of NaPi type II mRNA and protein was recorded by molecular biological methods. High intestinal Pi transport capacity was already present on the first day of life. Within the first 3 wk of life there seemed to be a change in the type of Na+-dependent Pi transporter, and NaPi IIb was expressed increasingly up to the fifth month of life. Renal Na+-Pi transport capacity was also high at birth, and this was associated with high expression levels of NaPi IIa mRNA, indicating the important role of this transporter for renal Pi reabsorption. At weaning an increase in both intestinal and renal Na+-Pi transport balanced the increasing requirements for Pi to establish the endogenous Pi cycle. Salivary Pi concentration and parotid NaPi II mRNA rose markedly to guarantee an adequate Pi supply for rumen microbes. We concluded that the high demand for Pi in young goats was assured by high basal Na+-Pi transport capacity of small intestines and kidney expressed continuously during ontogenesis. 相似文献
978.
Lindstrom WM Malygin EG Ovechkina LG Zinoviev VV Reich NO 《Journal of molecular biology》2003,325(4):711-720
We show that the kinetic mechanism of the DNA (cytosine-N(4)-)-methyltransferase M.BamHI, which modifies the underlined cytosine (GGATCC), differs from cytosine C(5) methyltransferases, and is similar to that observed with adenine N(6) methyltransferases. This suggests that the obligate order of ternary complex assembly and disassembly depends on the type of methylation reaction. In contrast, the single-turnover rate of catalysis for M.BamHI (0.10s(-1)) is closer to the DNA (cytosine-C(5)-)-methyltransferases (0.14s(-1)) than the DNA (adenine-N(6)-)-methyltransferases (>200s(-1)). The nucleotide flipping transition dominates the single-turnover constant for adenine N(6) methyltransferases, and, since the disruption of the guanine-cytosine base-pair is essential for both types of cytosine DNA methyltransferases, this transition may be a common, rate-limiting step for methylation for these two enzyme subclasses. The similar overall rate of catalysis by M.BamHI and other DNA methyltransferases is consistent with a common rate-limiting catalytic step of product dissociation. Our analyses of M.BamHI provide functional insights into the relationship between the three different classes of DNA methyltransferases that complement both prior structural and evolutionary insights. 相似文献
979.
Hippler-Mreyen S Klare JP Wegener AA Seidel R Herrmann C Schmies G Nagel G Bamberg E Engelhard M 《Journal of molecular biology》2003,330(5):1203-1213
Sensory rhodopsin II, a repellent phototaxis receptor from Natronobacterium pharaonis (NpSRII) forms a tight complex with its cognate transducer (NpHtrII). Light excitation of the receptor triggers conformational changes in both proteins, thereby activating the cellular two-component signalling cascade. In membranes, the two proteins form a 2:2 complex, which dissociates to a 1:1 heterodimer in micelles. Complexed to the transducer sensory rhodopsin II is no longer capable of light-driven proton pumping. In order to elucidate the dimerisation and the size of the receptor-binding domain of the transducer, isothermal titration calorimetry and electrophysiological experiments have been carried out. It is shown, that an N-terminal sequence of 114 amino acid residues is sufficient for tight binding (K(d)=240nM; DeltaH=-17.6kJmol(-1)) and for inhibiting the proton transfer. These data and results obtained from selected site-directed mutants indicate a synergistic interplay of transducer transmembrane domain (1-82) and cytoplasmic peptide (83-114) leading to an optimal and specific interaction between receptor and transducer. 相似文献
980.
Different heparan sulfate proteoglycans serve as cellular receptors for human papillomaviruses 总被引:6,自引:0,他引:6
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Shafti-Keramat S Handisurya A Kriehuber E Meneguzzi G Slupetzky K Kirnbauer R 《Journal of virology》2003,77(24):13125-13135
Papillomaviruses replicate in stratified epithelia of skin and mucosa. Infection with certain human papillomavirus (HPV) types is the main cause of anogenital neoplasia, in particular cervical cancer. Early events of papillomavirus infectivity are poorly understood. While heparan sulfate proteoglycans (HSPGs) mediate initial binding to the cell surface, the class of proteins carrying heparan sulfates has not been defined. Here we examined two processes of papillomavirus infection, attachment of virus-like particles (VLP) to cells and infection with authentic HPV type 11 (HPV11) virions. Of the HSPGs, syndecan-1 is the major epithelial form and is strongly upregulated in wound edge keratinocytes. We employed K562 cells, which lack HSPGs except minor amounts of endogenous betaglycan, and stable clones that express cDNAs of syndecan-1, syndecan-4, or glypican-1. Binding of VLP correlated with levels of heparan sulfate on the cell surface. Parental K562 bound HPV16 VLP weakly, whereas all three K562 transfectants demonstrated enhanced binding, with the highest binding capacity observed for syndecan-1-transfected cells, which also expressed the most HSPG. For HPV11 infectivity assays, a high virion inoculum was required to infect K562 cells, whereas ectopic expression of syndecan-1 increased permissiveness eightfold and expression of syndecan-4 or glypican-1 fourfold. Infection of keratinocytes was eliminated by treatment with heparitinase, but not phospholipase C, further implicating the syndecan family of integral membrane proteins as receptor proteins. Human keratinocytes with a homozygous deletion of alpha6 integrin are permissive for HPV11 infection. These results indicate that several HSPGs can serve as HPV receptors and support a putative role for syndecan-1, rather than alpha6 integrin, as a primary receptor protein in natural HPV infection of keratinocytes. 相似文献