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31.
New insights in Salicornia L. and allied genera (Chenopodiaceae) inferred from nrDNA sequence data 总被引:1,自引:0,他引:1
A phylogenetic analysis was performed based on ITS DNA sequences of fourteen samples from different sources of six species of Salicornia, the three allied genera Arthrocnemum, Sarcocornia and Halocnemum of the same tribe Salicornieae, and other genera of the subfamily Salicornioideae used in previous studies. Bassia hirsuta, Camphorosma monspeliaca (subfamily Chenopodioideae) and four species of Suaeda (subf. Suaedoideae) were chosen as outgroups. Results show that the annual genus Salicornia is a sister group to the perennial genera Sarcocornia, Arthrocnemum and Halocnemum. Moreover, the phylogenetic analysis based on ITS results distinguished two groups of Salicornia species which fitted with ploidy level: one group consisted of diploid species, and the second of tetraploid ones. Sarcocornia and Arthrocnemum are shown to be closely related, even though the species investigated here exhibited an evident distance between their ITS sequences. On the basis of our results, these two genera should be united. Bienertia (already separated as Bienertieae) was confirmed as probable outgroup to the subf. Salicornioideae, while Kalidium (subf. Salicornioideae, tribe Halopeplideae) was an outgroup to the rest of the Salicornioideae (tribe Salicornieae). The group Allenrolfea plus Halocnemum was the most basal of the tribe Salicornieae amongst those investigated in this study. The two samples of Halocnemum strobilaceum used in this work displayed numerous changes (transitions and transversions) in their respective sequences, probably related to their morphological and chorological differentiation. On the basis of our analysis, the most probable basal chromosome number for Salicornieae appears to be 2n = 18. The same number would also be the base number for the annual genus Salicornia and the perennial Arthrocnemum ( + Sarcocornia), with polyploidy arising independently in the two groups. 相似文献
32.
Substructure of the glomerular slit diaphragm in freeze-fractured normal rat kidney 总被引:4,自引:3,他引:1
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In the renal glomerulus, the narrow slits between adjacent epithelial podocytes are bridged by a diaphragm (2, 8, 11). In rat and mouse kidneys fixed by perfusion with tannic acid and glutaraldehyde (TAG), it has recently been discovered that this diaphragm has a highly ordered, isoporous substructure (9). It consists of a regular array of alternating cross bridges extending from the podocyte plasma membranes to a centrally running filament. This zipperlike pattern results in two rows of rectangular pores, approximately 40 X 140 A in cross section, dimensions consistent with the proposed role of the diaphragm as an important filtration barrier to plasma proteins (6). In the present study, we found in freeze-cleaved and in freeze-etched normal rat glomeruli that the surface of the slit diaphragm has an appearance conforming to the pattern found in sectioned material. 相似文献
33.
Solvent isotope effects and the pH dependence of laccase catalysis under steady-state conditions were examined with a rapid reductant to assess the potential roles of protein protic groups and the catalytic mechanism. The pH dependence of both reductant-dependent and reductant-independent steps showed bell-shaped profiles implicating at least two protic groups in each case. The apparent pKa values were: for the reductant-independent step(s), pK alpha 1 = 8.98 +/- 0.02 and pK alpha 2 = 5.91 +/- 0.03; for the reductant-dependent step(s), pK' alpha 1 = 7.55 +/- 0.12, pK' alpha 2 = 8.40 +/- 0.23. No solvent isotope effect on reductant-dependent steps was detected other than a standard shift effect. However, a significant solvent isotope effect on a reductant-independent step(s) was observed; kH/kD = 2.12 at the pH optimum of 7.5. The concentration dependence of the D2O effect indicated that a single proton was involved. Simulations of the p(H,D) data suggested that the solvent isotope effect was associated with the protein protic group required in its undissociated form (pK alpha 2). The pH effects on reductant-dependent steps are apparently associated with reductant-dependent steps that occur between O2 binding and water formation in the catalytic reaction sequence. 相似文献
34.
The bacteriophage 434 repressor distinguishes between its six naturally occurring binding sites using indirect readout. In indirect readout, sequence-dependent differences in the structure and flexibility of non-contacted bases in a protein's DNA-binding site modulate the affinity of DNA for protein. The conformation and flexibility of a DNA sequence can be influenced by the interaction of the DNA bases or backbone with solution components. We examined the effect of changing the cation-type present in solution on the stability and structure of 434 repressor complexes with wild-type and mutant OR1 and OR3, binding sites that differ in their contacted and non-contacted base sequences. We find that the affinity of repressor for OR1, but not for OR3, depends remarkably on the type and concentration of monovalent cation. Moreover, the formation of a stable, specific repressor-OR1 complex requires the presence of monovalent cations; however, repressor-OR3 complex formation has no such requirement. Changing monovalent cation type alters the ability of repressor to protect OR1, but not OR3, from *OH radical cleavage. Altering the relative length of the poly(dA) x poly(dT) tract in the non-contacted regions of the OR1 and OR3 can reverse the cation sensitivity of repressor's affinities for these two sites. Taken together these findings show that cation-dependent alterations in DNA structure underlies indirect readout of DNA sequence by 434 repressor and perhaps other proteins. 相似文献
35.
36.
Expression, purification, and functional characterization of the carboxyl-terminal domain fragment of bacteriophage 434 repressor. 总被引:2,自引:2,他引:0
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The repressor protein of bacteriophage 434 binds to DNA as a dimer of identical subunits. Its strong dimerization is mediated by the carboxyl-terminal domain. Cooperative interactions between the C-terminal domains of two repressor dimers bound at adjacent sites can stabilize protein-DNA complexes formed with low-affinity binding sites. We have constructed a plasmid, pCT1, which directs the overproduction of the carboxyl-terminal domain of 434 repressor. The protein encoded by this plasmid is called CT-1. Cells transformed with pCT1 are unable to be lysogenized by wild-type 434 phage, whereas control cells are lysogenized at an efficiency of 1 to 5%. The CT-1-mediated interference with lysogen formation presumably results from formation of heteromeric complexes between the phage-encoded repressor and the plasmid-encoded carboxyl-terminal domain fragment. These heteromers are unable to bind DNA and thereby inhibit the repressor's activity in promoting lysogen formation. Two lines of evidence support this conclusion. First, DNase I footprinting experiments show that at a 2:1 ratio of CT-1 to intact 434 repressor, purified CT-1 protein prevents the formation of complexes between 434 repressor and its OR1 binding site. Second, cross-linking experiments reveal that only a specific heterodimeric complex forms between CT-1 and intact 434 repressor. This latter observation indicates that CT-1 interferes with 434 repressor-operator complex formation by preventing dimerization and not by altering the conformation of the DNA-bound repressor dimer. Our other evidence is also consistent with this suggestion. We have used deletion analysis in an attempt to define the region which mediates the 434 repressor-CT-1 interaction. CT-1 proteins which have more than the last 14 amino acids removed are unable to interfere with 434 repressor action in vivo. 相似文献
37.
Determinants that govern the recognition and uptake of Escherichia coli O157 : H7 by Acanthamoeba castellanii
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Predation by phagocytic predators is a major source of bacterial mortality. The first steps in protozoan predation are recognition and consumption of their bacterial prey. However, the precise mechanisms governing prey recognition and phagocytosis by protists, and the identities of the molecular and cellular factors involved in these processes are, as yet, ill‐characterized. Here, we show that that the ability of the phagocytic bacterivorous amoebae, Acanthamoeba castellanii, to recognize and internalize Escherichia coli, a bacterial prey, varies with LPS structure and composition. The presence of an O‐antigen carbohydrate is not required for uptake of E. coli by A. castellanii. However, O1‐antigen types, not O157 O‐antigen types, inhibit recognition and uptake of bacteria by amoeba. This finding implies that O‐antigen may function as an antipredator defence molecule. Recognition and uptake of E. coli by A. castellanii is mediated by the interaction of mannose‐binding protein located on amoebae's surface with LPS carbohydrate. Phagocytic mammalian cells also use mannose‐binding lectins to recognize and/or mediate phagocytosis of E. coli. Nonetheless, A. castellanii's mannose binding protein apparently displays no sequence similarity with any known metazoan mannose binding protein. Hence, the similarity in bacterial recognition mechanisms of amoebae and mammalian phagocytes may be a result of convergent evolution. 相似文献
38.
39.
Matthew N Davies Channa K Hattotuwagama David S Moss Michael GB Drew Darren R Flower 《BMC structural biology》2006,6(1):5-13
Background
MHC Class I molecules present antigenic peptides to cytotoxic T cells, which forms an integral part of the adaptive immune response. Peptides are bound within a groove formed by the MHC heavy chain. Previous approaches to MHC Class I-peptide binding prediction have largely concentrated on the peptide anchor residues located at the P2 and C-terminus positions. 相似文献40.
Julia Katharina Keppler Tomas Koudelka Kalpana Palani Mayra Christina Stuhldreier Friedrich Temps Andreas Tholey 《Journal of biomolecular structure & dynamics》2013,31(7):1103-1117
Reversible binding of small compounds through hydrophobic interactions or hydrogen bonding to food proteins (e.g. milk proteins) is a thoroughly researched topic. In contrast, covalent interactions are not well characterized. Here, we report a rare form of positive-cooperativity-linear binding of allyl isothiocyanate with β-lactoglobulin, resulting in the cleavage of a disulfide bond of the protein. We compared three methods (i.e. fluorescence quenching, equilibrium dialysis, and headspace–water equilibrium) to characterize the binding kinetics and investigated the molecular binding by mass spectrometry. The methodologies used were found to be comparable and reproducible in the presence of high and low ligand concentrations for fluorescence quenching and equilibrium-based methods respectively. 相似文献