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991.
Monophyly of primary photosynthetic eukaryotes: green plants, red algae, and glaucophytes 总被引:4,自引:0,他引:4
Rodríguez-Ezpeleta N Brinkmann H Burey SC Roure B Burger G Löffelhardt W Bohnert HJ Philippe H Lang BF 《Current biology : CB》2005,15(14):1325-1330
Between 1 and 1.5 billion years ago, eukaryotic organisms acquired the ability to convert light into chemical energy through endosymbiosis with a Cyanobacterium (e.g.,). This event gave rise to "primary" plastids, which are present in green plants, red algae, and glaucophytes ("Plantae" sensu Cavalier-Smith). The widely accepted view that primary plastids arose only once implies two predictions: (1) all plastids form a monophyletic group, as do (2) primary photosynthetic eukaryotes. Nonetheless, unequivocal support for both predictions is lacking (e.g.,). In this report, we present two phylogenomic analyses, with 50 genes from 16 plastid and 15 cyanobacterial genomes and with 143 nuclear genes from 34 eukaryotic species, respectively. The nuclear dataset includes new sequences from glaucophytes, the less-studied group of primary photosynthetic eukaryotes. We find significant support for both predictions. Taken together, our analyses provide the first strong support for a single endosymbiotic event that gave rise to primary photosynthetic eukaryotes, the Plantae. Because our dataset does not cover the entire eukaryotic diversity (but only four of six major groups in), further testing of the monophyly of Plantae should include representatives from eukaryotic lineages for which currently insufficient sequence information is available. 相似文献
992.
993.
Halberg F Cornélissen G Wall D Otsuka K Halberg J Katinas G Watanabe Y Halhuber M Bohn TM Delmore P Siegelova J Homolka P Fiser B Dusek J Sánchez de la Peña S Maggioni C Delyukov A Gorgo Y Gubin D Carandente F Schaffer E Rhodus N Borer K Sonkowsky RP Schwartzkopff O 《Biomedical instrumentation & technology / Association for the Advancement of Medical Instrumentation》2002,36(3):183-197
994.
An antibody to the putative aphid recognition site on cucumber mosaic virus recognizes pentons but not hexons 下载免费PDF全文
Bowman VD Chase ES Franz AW Chipman PR Zhang X Perry KL Baker TS Smith TJ 《Journal of virology》2002,76(23):12250-12258
Cucumber mosaic virus (CMV), the type member of the genus Cucumovirus (family Bromoviridae), is transmitted by aphids in a nonpersistent manner. Mutagenesis experiments identified the betaH-betaI loop of the capsid subunit as a potential key motif responsible for interactions with the insect vector. To further examine the functional characteristics of this motif, we generated monoclonal antibodies that bound to native virions but not to betaH-betaI mutants. Fab fragments from these antibodies were complexed with wild-type CMV and the virus-Fab structure was determined to 12-A resolution by using electron cryomicroscopy and image reconstruction techniques. The electron density attributed to the bound antibody has a turret-like appearance and protrudes from each of the 12 fivefold axes of the icosahedral virus. Thus, the antibody binds only to the pentameric clusters (pentons) of A subunits of the T=3 quasisymmetric virus and does not appear to bind to any of the B and C subunits that occur as hexameric clusters (hexons) at the threefold (quasi-sixfold) axes. Modeling and electron density comparisons were used to analyze the paratope-epitope interface and demonstrated that the antibody binds to three betaH-betaI loops in three adjacent A subunits in each penton. This antibody can discriminate between A and B/C subunits even though the betaH-betaI loop adopts the same structure in all 180 capsid subunits and is therefore recognizing differences in subunit arrangements. Antibodies with such character have potential use as probes of viral assembly. Our results may provide an additional rationale for designing synthetic vaccines by using symmetrical viral particles. 相似文献
995.
In previous work, we had identified stabilized forms of the cold-shock protein Bs-CspB from Bacillus subtilis in a combinatorial library by an in vitro selection procedure. In this library, the sequence positions 2, 3, 46, 64, 66, and 67 had been randomized, because Bs-CspB differs from the naturally thermostable homolog Bc-Csp from Bacillus caldolyticus, among others, at these six positions. For the most stable selected variant, the midpoint of thermal unfolding (tM) increased by 28.2 deg. C and the Gibbs free energy of unfolding (deltaG(D)) by 19 kJ/mol. Here, we analyzed by site-directed mutagenesis how the selected residues contribute individually to this strong stabilization. Val3 and Val66, which replace Glu3 and Glu66 of wild-type Bs-CspB, each contribute about 7 kJ/mol to stability, the Thr64Arg substitution contributes 4.5 kJ/mol, and 3.2 kJ/mol originate from the Ala46Leu replacement. Gly67 at the carboxy terminus is unimportant for stability, the Arg selected at position 2 is overall slightly destabilizing but improves the coulombic interactions. The best variant differs from Bc-Csp at all six positions; nevertheless, natural and in vitro selection followed similar principles. In both cases, negatively charged residues at the adjacent positions 3 and 66 are avoided, and a positively charged residue is introduced into this area of the protein surface. Its exact location is unimportant. It can be at position 3, as in the thermophilic Bc-Csp, or at positions 2 or 64, as in the most stable selected variant. These positively charged residues contribute to stability not by engaging in pairwise coulombic interactions with a specific carboxyl group, but by generally improving the charge distribution in this particular region of the protein surface. These coulombic effects contribute significantly to the thermostability of the cold-shock proteins. They are only weakly interdependent and best explained by the presence of a flexible ion network at the protein surface. Our results emphasize that surface positions are very good candidates for optimizing protein stability. 相似文献
996.
Continuum electrostatic models are used to examine in detail the mechanism of protein stabilization and destabilization due to salt near physiological concentrations. Three wild-type cold shock proteins taken from mesophilic, thermophilic, and hyperthermophilic bacteria are studied using these methods. The model is validated by comparison with experimental data collected for these proteins. In addition, a number of single point mutants and three designed sequences are examined. The results from this study demonstrate that the sensitivity of protein stability toward salt is correlated with thermostability in the cold shock protein family. The calculations indicate that the mesophile is stabilized by the presence of salt while the thermophile and hyperthermophile are destabilized. A decomposition of the salt influence at a residue level permits identification of regions of the protein sequences that contribute toward the observed salt-dependent stability. This model is used to rationalize the effect of various point mutations with regard to sensitivity toward salt. Finally, it is demonstrated that designed cold shock protein variants exhibit electrostatic properties similar to the natural thermophilic and hyperthermophilic proteins. 相似文献
997.
Host-guest principles are put into action advantageously by cyclodextrins that give remarkable sensor responses to halogenated hydrocarbons on mass-sensitive devices such as QMB (quartz micro balance) and SAW (surface acoustic wave) resonators. Modifications of the structure can tune the compound to different analytes and molecular modeling allows us to understand and predict specific host-guest interactions. The FT-IR analysis of CDCl3, incorporated into a partially methylated cyclodextrine, yields an astonishingly strong band shift of about 50 cm-1 to lower wave numbers, in contrast to the permethylated product that shows a displaced band of 10 cm-1 for the C-D stretching vibration. The explanation can be given by semiempirical methods and force field calculations. Two CDCl3 binding sites are revealed, one being a multicentered inclusion ([(n)\tilde] ~ 2195 cm-1{\tilde\nu}\sim 2195\;cm^{-1} of CDCl3 at the upper rim of the #-cyclodextrine cone, whereas the other is incorporated within the cavity ([(n)\tilde] ~ 2240 cm-1{\tilde\nu}\sim 2240\;cm^{-1}. 相似文献
998.
The photochemical behaviour of an analogous bacteriorhodopsin (9,12-Ph-BR) which contains the sterically fixed 9,12-phenylretinal has been investigated with picosecond spectroscopy. The following results have been obtained. No ground-state intermediate photoproduct is found in agreement with the previous observation that 9,12-Ph-BR does not exhibit proton pumping under illumination. The excited singlet state has a lifetime of τS = 10 ± 2 ps. This lifetime agrees favourably with the value calculated from the radiative lifetime τrad = 6.2 ns and the fluorescence quantum efficiency of 1.2·10−3. Excited-state absorption occurs which results in fluorescence in the ultraviolet region. These various observations differ drastically from the corresponding findings on bacteriorhodopsin. Most important for an understanding of the differences is the fact that 9,12-phenylretinal does not isomerize in the protein's binding site in contrast to retinal. Our data therefore suggest that the formation of the intermediate K observed in bacteriorhodopsin is accompanied by the all-trans to 13-cis isomerization. 相似文献
999.
1000.