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1.
Breydo L Bocharova OV Makarava N Salnikov VV Anderson M Baskakov IV 《Biochemistry》2005,44(47):15534-15543
In recent studies, we developed a protocol for in vitro conversion of full-length mouse recombinant PrP (Mo rPrP23-230) into amyloid fibrils [Bocharova et al. (2005) J. Mol. Biol. 346, 645-659]. Because amyloid fibrils produced from recombinant Mo PrP89-230 display infectivity [Legname et al. (2004) Science 305, 673-676], polymerizatiom of rPrPs in vitro represents a valuable model for elucidating the mechanism of prion conversion. Unexpectedly, when the same conversion protocol was used for hamster (Ha) rPrP23-231, we experienced substantial difficulties in forming fibrils. While searching for potential reasons of our failure to produce fibrils, we probed the effect of methionine oxidation in rPrP. We found that oxidation of methionines interferes with the formation of rPrP fibrils and that this effect is more profound for Ha than for Mo rPrP. To minimize the level of spontaneous oxidation, we developed a new protocol for rPrP purification, in which highly amyloidogenic Ha rPrP with minimal levels of oxidized residues was produced. Furthermore, our studies revealed that oxidation of methionines in preformed fibrils inhibited subsequent maturation of fibrils into proteinase K-resistant PrP(Sc)-like conformation (PrP-res). Our data are consistent with the proposition that conformational changes within the central region of the protein (residues 90-140) are essential for adopting PrP-res conformation and demonstrate that methionine oxidation interferes with this process. These studies provide new insight into the mechanism of prion polymerization, solve a long-standing practical problem in producing PrP-res fibrils from full-length PrP, and may help in identifying new genetic and environmental factors that modulate prion disease. 相似文献
2.
Bocharova OV Breydo L Salnikov VV Gill AC Baskakov IV 《Protein science : a publication of the Protein Society》2005,14(5):1222-1232
In recent studies, the amyloid form of recombinant prion protein (PrP) encompassing residues 89-230 (rPrP 89-230) produced in vitro induced transmissible prion disease in mice. These studies showed that unlike "classical" PrP(Sc) produced in vivo, the amyloid fibrils generated in vitro were more proteinase-K sensitive. Here we demonstrate that the amyloid form contains a proteinase K-resistant core composed only of residues 152/153-230 and 162-230. The PK-resistant fragments of the amyloid form are similar to those observed upon PK digestion of a minor subpopulation of PrP(Sc) recently identified in patients with sporadic Creutzfeldt-Jakob disease (CJD). Remarkably, this core is sufficient for self-propagating activity in vitro and preserves a beta-sheet-rich fibrillar structure. Full-length recombinant PrP 23-230, however, generates two subpopulations of amyloid in vitro: One is similar to the minor subpopulation of PrP(Sc), and the other to classical PrP(Sc). Since no cellular factors or templates were used for generation of the amyloid fibrils in vitro, we speculate that formation of the subpopulation of PrP(Sc) with a short PK-resistant C-terminal region reflects an intrinsic property of PrP rather than the influence of cellular environments and/or cofactors. Our work significantly increases our understanding of the biochemical nature of prion infectious agents and provides a fundamental insight into the mechanisms of prions biogenesis. 相似文献
3.
Ostrik A. A. Grigorov A. S. Bocharova I. V. Kaprelyants A. S. Azhikina T. L. Salina E. G. 《Applied Biochemistry and Microbiology》2022,58(4):401-405
Applied Biochemistry and Microbiology - The role of small non-coding RNAs Mcr11 and DrrS with its possible synergy in the metabolism of M. tuberculosis was studied. There were no noticeable... 相似文献
4.
本文用脑室灌注和Fura2测定细胞内游离钙技术观察了地塞米松(dexamethasone,DEX)对家兔乙二醇双(2氨基乙醚)四乙酸性发热效应和下丘脑细胞内游离钙浓度([Ca2+]i)的影响,借此深入探讨地塞米松解热作用的中枢机制。结果发现:脑室灌注乙二醇双(2氨基乙醚)四乙酸(06nmol)引起家兔结肠温度明显升高,静脉注射地塞米松(5mg/kg)显著抑制家兔乙二醇双(2氨基乙醚)四乙酸性发热,地塞米松(60~120μmol/L)并不影响下丘脑细胞内[Ca2+]i,而事先脑室灌注抑制基因转录的放线菌素D(3nmol)则完全取消了地塞米松对乙二醇双(2氨基乙醚)四乙酸性发热的解热作用。这些结果提示:地塞米松显著抑制家兔乙二醇双(2氨基乙醚)四乙酸性发热,其机制与地塞米松激活脑内某些基因的表达有关,而与下丘脑神经细胞跨膜钙离子流无关。 相似文献
5.
Fushitani K; Higashiyama K; Moriyama EN; Imai K; Hosokawa K 《Molecular biology and evolution》1996,13(7):1039-1043
To elucidate phylogenetic relationships among amniotes and the evolution of
alpha globins, hemoglobins were analyzed from the Komodo dragon (Komodo
monitor lizard) Varanus komodoensis, the world's largest extant lizard,
inhabiting Komodo Islands, Indonesia. Four unique globin chains (alpha A,
alpha D, beta B, and beta C) were isolated in an equal molar ratio by high
performance liquid chromatography from the hemolysate. The amino acid
sequences of two alpha chains were determined. The alpha D chain has a
glutamine at E7 as does an alpha chain of a snake, Liophis miliaris, but
the alpha A chain has a histidine at E7 like the majority of hemoglobins.
Phylogenetic analyses of 19 globins including two alpha chains of Komodo
dragon and ones from representative amniotes showed the following results:
(1) The a chains of squamates (snakes and lizards), which have a glutamine
at E7, are clustered with the embryonic alpha globin family, which
typically includes the alpha D chain from birds; (2) birds form a sister
group with other reptiles but not with mammals; (3) the genes for embryonic
and adult types of alpha globins were possibly produced by duplication of
the ancestral alpha gene before ancestral amniotes diverged, indicating
that each of the present amniotes might carry descendants of the two types
of alpha globin genes; (4) squamates first split off from the ancestor of
other reptiles and birds.
相似文献
6.
7.
Summary Several types of receptor endings were identified with scanning electron microscopy and silver-impregnation techniques in the skin of the tentacles, lips, dorsal surface of the head and mouth region of the pond snails Lymnaea stagnalis and Vivipara viviparus. Sensory endings at the tips of dendrites of primary receptor neurones, scattered below the epithelium, differ in structure, i.e., the endings exposed to the surface of the skin possess different proportions of cilia and microvilli, which vary in number, length, and packing. Type-I endings have microvilli and a few (1–5) cilia, 5–12 m in length. Type-2 endings have abundant (20–40), interwoven long (9–12 m) cilia and random microvilli. Type-3 endings show typical packing of 10–25 cilia in the form of bundles or brushes. They may be composed either of long (9–18 m) or short (2–7 m) cilia, or of both long and short ones. Microvilli here are absent. Type-4 endings have only microvilli. Two other types of skin receptors do not extend their sensory endings to the surface and can be indentified only in silver-stained preparations. Type-5 endings are branching dendrites of skin receptors cells that terminate among epithelial cells. In type-6, the sensory endings also terminate among epithelial cells but their cell bodies are located outside of the skin. In both species all skin regions examined possess the receptors of all six types differing only in their relative proportion. Possible functional roles of different receptors are discussed. 相似文献
8.
V. D. Lazareva N. E. Bocharova K. E. Volynski T. M. Volkova E. V. Grishin 《Russian Journal of Bioorganic Chemistry》2000,26(4):271-276
α-Latrotoxin (α-LTX) binding sites to functionally active monoclonal antibodies (MA) A4 and A24 were localized using three
approaches: hydrolysis of the toxin followed by theN-terminal sequencing of immunoreactive peptides; the study of antibody interaction with several recombinant α-LTX fragments;
Western immunoblotting of synthetic overlapping peptides (6–8 aa) whose structures correspond to that of the immunoreactive
α-LTX fragment. It was shown that the MA A4 epitope is located within the F234-M294 protein fragment and that MA A24 interacts with the fragment347FDKDIT352. 相似文献
9.
Landscape-scale variation in the seed banks of floodplain wetlands with contrasting hydrology in China 总被引:3,自引:0,他引:3
1. At a local scale, the species composition, diversity and spatial variation of wetland plant communities are determined primarily by spatial and temporal heterogeneity in their environments. Less is known about variation at a landscape‐level. The floodplain of the Changjiang (Yangtze) River in China includes hydrologically connected, subtropical wetlands with different hydrological characteristics. 2. We examined seed‐bank species composition and richness in marshes of two contrasting hydrological types: permanent marshes, fed by local runoff, and lakeshore marshes more closely connected to the regulated river. Lakeshore marshes are flooded annually to depth of approximately 1 m and during flooding they support an alternate, aquatic vegetation type. The soil seed bank in March was a comparative estimator of species diversity. At the beginning of the growing season it included seeds from both phases of alternating vegetation types associated with the annual hydrological cycle. 3. A regional pool of 101 species was detected in the seed banks of six wetlands associated with the river and its tributaries: 56 occurred in permanent marshes and 59 in lakeshore marshes, with only 15 common to both. Species rarefaction curves indicated that more species occurred in permanent than lakeshore marshes at equal numbers of individuals sampled. However, the more heterogeneous lakeshore seed banks were estimated (Chao 2) to have greater total species richness (81) than permanent marsh (60). 4. Analysis using Sørensen's coefficient of similarity and DCA ordination revealed complex variation, with much greater differences between hydrological types than within them, irrespective of geographical distance. The types also differed significantly in the composition of four functional groups of species. 5. Despite the potential for dispersal of propagules via the annually pulsing river system (hydrochory), at a regional and landscape scale, diversity is maintained largely by large‐scale temporal hydrological heterogeneity and smaller scale spatial and topographic heterogeneity. 相似文献
10.