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91.
The crystal structure of glycerol-3-phosphate cytidylyltransferase from B. subtilis (TagD) is about to be solved. Here, we report a testable structure prediction based on the identification by sequence analysis of a superfamily of functionally diverse but structurally similar nucleotide-binding enzymes. We predict that TagD is a member of this family. The most conserved region in this superfamily resembles the ATP-binding HiGH motif of class I aminoacyI-tRNA synthetases. The predicted secondary structure of cytidylyltransferase and its homologues is compatible with the α/β topography of the class I aminoacyl-tRNA synthetases. The hypothesis of similarity of fold is strengthened by sequence-structure alignment and 3D model building using the known structure of tyrosyl tRNA synthetase as template. The proposed 3D model of TagD is plausible both structurally, with a well packed hydrophobic core, and functionally, as the most conserved residues cluster around the putative nucleotide binding site. If correct, the model would imply a very ancient evolutionary link between class I tRNA synthetases and the novel cytidylyltransferase superfamily. © 1995 Wiley-Liss, Inc. 相似文献
92.
Dmitry E. Nolde Alexander G. Sobol Kirill A. Pluzhnikov Eugene V. Grishin Alexander S. Arseniev 《Journal of biomolecular NMR》1995,5(1):1-13
Summary Two-dimensional 1H NMR techniques were used to determine the spatial structure of ectatomin, a toxin from the venom of the ant Ectatomma tuberculatum. Nearly complete proton resonance assignments for two chains of ectatomin (37 and 34 amino acid residues, respectively) were
obtained using 2D TOCSY, DQF-COSY and NOESY experiments. The cross-peak volumes in NOESY spectra were used to define the local
structure of the protein and generate accurate proton-proton distance constraints employing the MARDIGRAS program. Disulfide
bonds were located by analyzing the global fold of ectatomin, calculated with the distance geometry program DIANA. These data,
combined with data on the rate of exchange of amide protons with deuterium, were used to obtain a final set of 20 structures
by DIANA. These structures were refined by unrestrained energy minimization using the CHARMm program. The resulting rms deviations
over 20 structures (excluding the mobile N- and C-termini of each chain) are 0.75 ? for backbone heavy atoms, and 1.25 ? for
all heavy atoms. The conformations of the two chains are similar. Each chain consists of two α-helices and a hinge region
of four residues; this forms a hairpin structure which is stabilized by disulfide bridges. The hinge regions of the two chains
are connected together by a third disulfide bridge. Thus, ectatomin forms a four-α-helical bundle structure. 相似文献
93.
Plasmodium falciparum protein associated with the invasion junction contains a conserved oxidoreductase domain 总被引:2,自引:0,他引:2
Diana E. Hudson-Taylor Stephen A. Dolan Francis W. Klotz Hisashi Fujioka Masamichi Aikawa Eugene V. Koonin Louis H. Miller 《Molecular microbiology》1995,15(3):463-471
The merozoite cap protein-1 (MCP-1) of Plasmodium falciparum follows the distribution of the moving Junction during invasion of erythrocytes. We have cloned the gene encoding this protein from a cDNA library using a monoclonal antibody. The protein lacks a signal sequence and has no predicted trans-membrane domains; none of the antisera reacts with the surfaces of intact merozoites, indicating that the cap distribution is submembranous. MCP-1 is divided into three domains. The N-terminal domain includes a 52-amino-acid region that is highly conserved in a large family of bacterial and eukaryotic proteins. Based on the known functions of two proteins of this family and the pattern of amino acid conservation, it is predicted that this domain may possess oxido-reductase activity, since the active cysteine residue of this domain is invariant in all proteins of the family. The other two domains of MCP-1 are not found in any other members of this protein family and may reflect the specific function of MCP-1 in invasion. The middle domain is negatively charged and enriched in glutamate; the C-terminal domain is positively charged and enriched in lysine. By virtue of its positive charge, the C-terminal domain resembles domains in some cytoskeleton-associated proteins and may mediate the interaction of MCP-1 with cytoskeleton in Plasmodium. 相似文献
94.
Huckleberry plants evaluated at 13 sites in the southern Cascade Mountains of Washington State during August 1980 showed significantly lower fruit yields where subjected to heavy ash deposition following the 18 May 1980 eruption of Mt. St. Helens. High insect pollinator mortality is suspected as the major causal factor. The impact of such effects of volcanic activity on Native American subsistence is discussed. 相似文献
95.
Antibodies to synthetic myelin basic protein peptide S82 (TTHYG-SLPQKAQGHRPQDEG) did not react with synthetic peptide S8 (GSLPQKAQGHRPQDENG) and only partially so with synthetic peptide S79 (AQGHRPQDEG); however, the antibodies did react to a considerable extent with an equimolar mixture of S8 and S79. Since the anti-S82 antibodies had previously been shown to be directed to a non-sequential format determinant dependent on the conformation of secondary structure, it seems probable that the mixture of S8 and S79 assumed a format that neither one individually possessed to any great degree.Special Issue dedicated to Dr. Elizabeth Roboz-Einstein.Supported by Research Grants NS-10237 (Duke) and NS-15322 (St. Luke's) from the National Institutes of Health and by RG1197-B7 from the National Multiple Sclerosis Society. 相似文献
96.
Compared to littermate controls, unstressed Jimpy mice have higher brain water, sodium, potassium and chloride contents and lower carbonic anhydrase activity. When stressed by CO2 to produce a respiratory acidosis or by injection of distilled water to produce brain edema, the Jimpy mouse brain has water and ionic responses essentially like those in controls. 相似文献
97.
Isolation of a recombinant lambda phage carrying nusA and surrounding region of the Escherichia coli K-12 chromosome 总被引:5,自引:0,他引:5
Summary A recombinant bacteriophage lambda, argG-6, has been isolated which carries the argG gene and neighbouring loci on an EcoRI-generated 15.5 Kb DNA fragment from the Escherichia coli chromosome. The locations of the argG, nusA and pnp genes on the 15.5 Kb DNA fragment have been determined. In the case of nusA, a Tn5 insertion and sub-cloning of restriction fragments were used to locate the gene. The gene products of nusA and pnp have been identified on one- and two-dimensional polyacrylamide gels. The clockwise gene order was found to be argG-nusA-pnp. 相似文献
98.
W. Scott Ramsey Eugene D. Nowlan Lynn B. Simpson 《Applied microbiology and biotechnology》1980,9(3):217-226
Summary The resolution of bacterial mixtures by free flow electrophoresis (FFE) was not affected by the position of the microbes on the growth curve and approximately 70% of the individual cells applied were recovered as viable cells. The dependence of bacterial electrophoretic mobility on the pH, salt concentration, and viscosity of the electrolyte was determined. Suspending media and running electrolyte were developed which allowed collection of samples of>99% purity within two minutes of introduction of a mixture of Escherichia coli and Staphylococcus aureus. Most bacterial strains migrated in a single band, although some migrated in more than one band. Escherichia coli was resolved from each of 10 different species. The considerable variation in mobility found in 21 different E. coli strains, however, appears to preclude use of FFE as a method of species identification. 相似文献
99.
Reduction of 3-ketosucrose (1) with sodium borohydride gave mainly α-d-allopyranosyl β-d-fructofuranoside (2) characterized as its octabenzoate. Using sodium borodeuteride, [3-2H]allo-sucrose (5) and [3-2H]sucrose (6) were obtained in the ratio 12:1. The mixture was fractionated on Dowex-50 X8 resin (Ca2+ form), and the [3-2H] derivatives were isolated as their octa-acetates. Inspection of the 13C-n.m.r. spectra of 5 and 6 enabled the C-3 signals to be assigned. allo-Sucrose (2) was more readily obtained by oxidation of sucrose with dimethyl sulphoxide-acetic anhydride followed by reduction with sodium borohydride and fractionation on Dowex-50 X8 (Ca2+) resin. Tritylation of 2 followed by acetylation gave, after chromatography, the 6,1′,6′-tritrityl ether (9, 10%), the 6,6′-ditrityl ether (10, 26%), and a mixture of monotrityl ethers (20%). Hydrogenolysis of 9 and 10 gave the penta-acetate and hexa-acetate, respectively, with no detectable migration of AcO-4. Treatment of 2 with sulphuryl chloride at -50° gave the 6,6′-dichloride. 相似文献
100.
Development of two cloned epithelial cell lines from normal adult mouse and rat ventral prostates 总被引:1,自引:0,他引:1
Yoshinobu Kubota Eugene B. Gehly Karl H. Link Charles Heidelberger 《In vitro cellular & developmental biology. Plant》1981,17(11):965-978
Summary Two epithelial cell lines were established, one from adult C3H mouse and one from adult Fischer rat ventral prostate. These
cell lines were obtained from explant cultures, using Ham's F12 medium supplemented with HEPES, insulin, testosterone, hydrocortisone,
epidermal growth factor, and 7.5% fetal bovine serum. A low concentration of trypsin and EDTA in Ca++-and Mg++-free phosphate buffer was used for passaging the cells. The rat cell line was established following implantation of prostate
tissue in nude mice. These cell lines stained positively for acid phosphatase and were dependent upon epidermal growth factor
for growth. Morphological studies, including electron microscopy, revealed a highly characteristic epithelial morphology of
both cell lines. These cell lines have hypotetraploid chromosome numbers and are capable of metabolizing benzo(a)pyrene. We propose the application of these cells as models for the study of prostate carcinogenesis.
This work was supported in part by Grant CA-21, 746, and by the Electron Microscope Core Facility on Grant CA-14,089, from
the National Cancer Institute, National Institutes of Health, Bethesda, MD. 相似文献