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101.
Hubert Wiener Dan A. Klaerke Peter L. Jørgensen 《The Journal of membrane biology》1990,117(3):275-283
Summary In the mammalian distal colon, the surface epithelium is responsible for electrolyte absorption, while the crypts are the site of secretion. This study examines the properties of electrical potential-driven86Rb+ fluxes through K+ channels in basolateral membrane vesicles of surface and crypt cells of the rabbit distal colon epithelium. We show that Ba2+-sensitive, Ca2+-activated K+ channels are present in both surface and crypt cell derived vesicles with half-maximal activation at 5×10–7
m free Ca2+. This suggests an important role of cytoplasmic Ca2+ in the regulation of the bidirectional ion fluxes in the colon epithelium.The properties of K+ channels in the surface cell membrane fraction differ from those of the channels in the crypt cell derived membranes. The peptide toxin apamin inhibits Ca2+-activated K+ channels exclusively in surface cell vesicles, while charybdotoxin inhibits predominantely in the crypt cell membrane fraction. Titrations with H+ and tetraethylammonium show that both high-and low-sensitive86Rb+ flux components are present in surface cell vesicles, while the high-sensitive component is absent in the crypt cell membrane fraction. The Ba2+-sensitive, Ca2+-activated K+ channels can be solubilized in CHAPS and reconstituted into phospholipid vesicles. This is an essential step for further characterization of channel properties and for identification of the channel proteins in purification procedures. 相似文献
102.
Multiple T and B cell epitopes in the S1 subunit ("A"-monomer) of the pertussis toxin molecule 总被引:5,自引:0,他引:5
J R Oksenberg C Ko A K Judd M Lim A Kent G K Schoolnik L Steinman 《Journal of immunology (Baltimore, Md. : 1950)》1989,143(12):4227-4231
The immunogenicity and reactogenicity of Bordetella pertussis vaccine are mediated in part by the S1 subunit of pertussis toxin (PT). To identify the immune epitopes in the S1 subunit of PT, synthetic peptides were prepared and tested for their capacity to induce antibodies in mice with different MHC genotypes. In BALB/c mice, peptides corresponding to sequences 1-17, 70-82 and 189-199 generate T cell proliferative responses, induce the production of antibodies capable of neutralization of the toxin in the Chinese hamster ovary-cell assay, and protect mice from a shock-like syndrome caused by alternate injections of BSA and PT. Protection and neutralization correlated with the ability of these peptides to elicit high anti-PT titers. Different B cell epitopes were detected in other inbred mouse strains. The antibody reactivity against synthetic peptides from two infants vaccinated with pertussis vaccine was tested. These infants had antibodies reactive to a variety of epitopes in the S1 subunit, including peptides 1-17, 70-82, 99-112, 135-145, and 189-199. Thus, it appears that there are multiple T and B cell epitopes in the S1 subunit of PT. 相似文献
103.
We have characterized the properties of a set of variants of the N-terminal domain of lambda repressor bearing disruptive mutations in the hydrophobic core. These mutations include some that dramatically alter the total core residue volume (by up to six methylene groups) and some that place a single polar residue into the otherwise hydrophobic core. The structural properties of the purified proteins have been studied by CD spectroscopy, biological activity, recognition by conformation-specific monoclonal antibodies, and 1H NMR spectroscopy. The stabilities of the proteins have been measured by thermal and guanidine hydrochloride denaturation. Proteins with disruptive core mutations are found to display a continuum of increasingly nonnative properties. Large internal volume changes cause both significant conformational rearrangements and destabilization by up to 5 kcal/mol. Variants with polar substitutions at core positions no longer behave like well-folded proteins but rather display characteristics of molten globules. However, even proteins bearing some of the most disruptive mutations retain many of the crude secondary and tertiary structural features of the wild-type protein. These results indicate that primitive elements of native structure can form in the absence of normal core packing. 相似文献
104.
昆虫是森林生态系统中的重要组成部分。由于森林中枯枝落叶较多,生境复杂,为昆虫提供了良好的繁殖和栖息场所;因而在森林中昆虫的种类多,数量大。大多数昆虫都以森林 相似文献
105.
David A. Raftos Dan L. Stillman Edwin L. Cooper 《In vitro cellular & developmental biology. Plant》1990,26(10):962-970
Summary Pharyngeal explants and circulatory hemocytes from the tunicateStyela clava were cultured in a medium containing tunicate plasma, artificial seawater, RPMI 1640, and antibiotics. Pharnngeal tissue
remained viable and proliferated for up to 72 d in vitro. Proliferative activity maintained the pool of hemocytes within explants
and facilitated the migration of pharyngeal hemocytes from explants into culture supernatants. The diversity of morphologically
distinct cell types within the hemocyte pool of pharyngeal cultures indicated that cell division was followed by regulated
differentiation. In contrast to pharyngeal cultures, suspensions of circulatory hemocytes did not survive for prolonged periods
in vitro. Proliferative activity could not be detected in circulatory hemocyte cultures. These results are discussed in terms
of the differentiation state of hemocytes and the efficacy of culture conditions.
This study was supported by the National Science Foundation, Washington, DC (grant DCB 85 19848) and by BRSG funds from UCLA
Schools of Medicine and Dentistry. Flow cytometric facilities were sponsored in part by a Johnson Cancer Center Core Grant
(CA 16042). David A. Raftos is a Fulbright Postdoctoral Fellow and recipient of a Frederik B. Bang Scholarship in Marine Invertebrate
Immunology administered by the American Association of Immunologist. Dan L. Stillman was supported by an REU supplement from
the National Science Foundation. 相似文献
106.
Recombinant human glia maturation factor beta (GMF-beta) reversibly inhibits the proliferation of neoplastic cells in culture by arresting the cells in the G0/G1 phase. This phenomenon is not target-cell specific, as neural and nonneural cells are equally inhibited. When tested simultaneously, GMF-beta suppresses the mitogenic effect of acidic fibroblasts growth factor (aFGF), but the two are synergistic in promoting the morphologic differentiation of cultured astrocytes. GMF-beta also counteracts the growth-stimulating effect of pituitary extract and cholera toxin on Schwann cells. The results underscore the regulatory role of GMF-beta and its intricate interaction with the mitogenic growth factors. 相似文献
107.
To help investigate the evolutionary origin of the imprinting (parent-of-origin mono-allelic expression) of paternal genes observed in mammals, we constructed haploid and diploid androgenetic zebrafish (Danio rerio). Haploid androgenotes were produced by fertilizing eggs that had been X-ray irradiated to eliminate the maternal genome. Subsequent inhibition of the first mitotic division of haploid androgenotes by heat shock produced diploid androgenotes. The lack of inheritance of maternal-specific DNA markers (RAPD and SSR) by putative diploid and haploid androgenotes confirmed the androgenetic origin of their genomes. Marker analysis was performed on 18 putative androgenotes (five diploids and 13 haploids) from six families. None of 157 maternal-specific RAPD markers analyzed, some of which were apparently homozygous, were passed on to any of these putative androgenotes. A mean of 7.7 maternal-specific markers were assessed per family. The survival of androgenetic zebrafish suggests that if paternal imprinting occurs in zebrafish, it does not result in essential genes being inactivated when their expression is required for development. Production of haploid androgenotes can be used to determine the meiotic recombination rate in male zebrafish. Androgenesis may also provide useful information about the mechanism of sex determination in zebrafish. 相似文献
108.
Helper virus-free transfer of herpes simplex virus type 1 plasmid vectors into neural cells. 总被引:11,自引:2,他引:9 下载免费PDF全文
C Fraefel S Song F Lim P Lang L Yu Y Wang P Wild A I Geller 《Journal of virology》1996,70(10):7190-7197
Herpes simplex virus type 1 (HSV-1) plasmid vectors have promise for genetic intervention in the brain, but several problems caused by the helper virus have compromised their utility. To develop a helper virus-free packaging system for these vectors, the DNA cleavage/packaging signals were deleted from a set of cosmids that represents the HSV-1 genome. Following cotransfection into cells, this modified cosmid set supported replication and packaging of vector DNA. However, in the absence of the DNA cleavage/packaging signals, the HSV-1 genome was not packaged, and consequently vector stocks were free of detectable helper virus. In the absence of helper virus, the vectors efficiently infected rat neural cells in culture or in the brain with minimal cytopathic effects. beta-galactosidase-positive cells were observed for at least 1 month in vivo, and vector DNA persisted for this period. This system may facilitate studies on neuronal physiology and potential therapeutic applications. 相似文献
109.
110.