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81.
Human neutrophil response to recombinant neisserial Opa proteins 总被引:13,自引:0,他引:13
Interactions of human neutrophils with recombinant Escherichia coli expressing gonococcal outer membrane Opa proteins were examined using chemiluminescent and biological assays. Seven opa loci from Neisseria gonorrhoeae MS11 4.8 were expressed as beta-lactamase-Opa fusion proteins that contained all but the mature N-terminal amino acid of the full-length Opa protein fused to three N-terminal amino acids derived from the mature beta-lactamase. The Opa fusion proteins were exported and assembled in the outer membrane of E. coli in a manner similar to that of Opa in N. gonorrhoeae, as evaluated by antibody binding and in situ proteolytic cleavage. All fusion proteins exhibited the characteristic heat-modifiable migration in SDS-polyacrylamide gel electrophoresis that typifies Opa proteins of neisseriae. Opa fusion proteins conferred on E. coli the ability to stimulate a chemiluminescent response from human neutrophils in the absence of antibody or complement. The nature of the response in terms of chemiluminescence, phagocytosis, and killing was in all cases analogous to that seen using N. gonorrhoeae expressing the equivalent Opa protein. Neither E. coli nor gonococci expressing OpaA elicited a response from neutrophils. Use of E. coli expressing Opa fusions should be useful in defining their biological activities and pathogenic roles. 相似文献
82.
83.
Ann E. Wakefield Sarah E. Peters Suneale Banerji Paul D. Bridge Geoffrey S. Hall David L. Hawksworth Lynden A. Guiver rew G. Allen Julian M. Hopkin 《Molecular microbiology》1992,6(14):1903-1911
Pneumocystis carinii causes life-threatening pneumonia in T-lymphocyte-immunodeficient subjects in transplant and oncology units or with acquired immune deficiency syndrome (AIDS). Recent DNA homology studies show P. carinii to be a fungus. To investigate the biology and epidemiology of this parasite further, we elected to determine for it a more precise taxonomic assignment within the fungal kingdom. We screened a wide range of organisms representing the major orders of fungi using DNA amplification and subsequently sequenced a portion of the mitochondrial gene encoding the large subunit ribosomal RNA. Our data show that the opportunistic pulmonary pathogen P. carinii is closely related to the ustomycetous red yeast fungi, a group which includes organisms that are extensively distributed throughout the environment and which release many widely dispersed airborne spores. 相似文献
84.
Identification and characterization of an {alpha}-mannosidase from Trypanosoma cruzi 总被引:1,自引:0,他引:1
Swanson Paul M.; Carter Clint E.; Hager Cindy; Kim Wan Joon; Obermeier Sarah; Oeltmann Thomas N. 《Glycobiology》1992,2(6):563-569
In this report we describe the first purification and characterizationof the acid -mannosidase from the human parasite Trypanosomacruzi. The purified enzyme exhibited a native mol. wt of 240000 Da and is apparently composed of four identical subunitsof mol. wt 58 000 Da. Each of the four subunits contains oneN-linked high-mannose-type oligosaccharide. The -mannosidaseexhibited a pH optimum of 3.5 and a pI of 5.9. This low pH optimumand the ability of swainsonine to inhibit its activity suggestthat the -mannosidase is a lysosomal enzyme. Antibodies againstthe T.cruzi enzyme did not react with mammalian lysosomal -mannosidaseand, conversely, antibody against a rat lysosomal -mannosidasedid not react with the T.cruzi enzyme. Thus, the T.cruzi enzymeappears to be distinct from its mammalian counterpart. -mannosidase lysosomal enzyme Trypanosoma cruzi 相似文献
85.
The adenine nucleotide translocator of higher plants is synthesized as a large precursor that is processed upon import into mitochondria. 总被引:6,自引:0,他引:6
B M Winning C J Sarah P E Purdue C D Day C J Leaver 《The Plant journal : for cell and molecular biology》1992,2(5):763-773
Two maize genes and cDNAs encoding the mitochondrial adenine nucleotide translocator (ANT), a nuclear-encoded inner mitochondrial membrane carrier protein, have previously been isolated in this laboratory. Sequence analysis revealed the existence of much longer open reading frames than the corresponding fungal and mammalian ANT genes. Potato ANT cDNAs have subsequently been isolated and sequenced and alignment of the deduced plant amino acid sequences with the equivalent fungal and mammalian polypeptides indicated that the plant proteins contain N-terminal extensions. When the plant cDNA clones are expressed in vitro they direct the synthesis of precursor proteins that are specifically processed at the N-terminus upon import into isolated mitochondria. N-terminal amino acid sequence data obtained from the native proteins purified from both maize and potato mitochondria has allowed identification of the putative processing sites. Further import analysis has shown that two distinct regions of the maize precursor protein contain targeting information, the 97 amino acids at the N-terminus and the 267 C-terminal amino acids. This is the first report that provides experimental evidence that the adenine nucleotide translocator of higher plants is synthesized as a large precursor protein that is specifically cleaved upon import into mitochondria. Import of ANT into higher plant mitochondria therefore appears to be different to the corresponding process in fungal and mammalian systems where targeting of ANT to mitochondria is mediated by internal signals and there is no N-terminal processing. 相似文献
86.
Sarah J. Holden D. Layne Coppock Mulugeta Assefa 《Human ecology: an interdisciplinary journal》1991,19(1):35-59
Surveys of pastoral households in a semi-nomadic Borana community during 1987–1988 were used to test the hypothesis that poorer families living closest to a market town would be most affected by the enhanced opportunity to sell dairy products, which would intensify competition between people and calves for milk and have negative implications for calf management. These poorer families indeed reported the highest rates of milk offtake per cow, and the milk increment was probably sold to purchase more grain for human consumption at the expense of milk intake for the calf. Consequently, this strategy may increase the susceptibility of malnourished calves to disease, especially those from lower-producing dams. Benefits of improved human energy intake from grain and retention of livestock capital must be weighed against risks of calf death and possible malnutrition of people from milk restriction when assessing dairy marketing trade-offs that are most acute for the poor. Opportunity to sell dairy products at favorable terms of trade helps the poorest people survive, and their risks could be mitigated by policies that facilitate grain marketing in the rangelands and interventions that improve calf feeding management, diversify human diets, and create alternative opportunities for women to generate income. The households postulated to be most at risk were identified from a complex, but logical, interaction among factors of distance to market, household wealth, and the quality of milking cows held. This indicates that targeting such needy groups for development assistance may require a more detailed and interdisciplinary analysis of production systems than is commonly practiced. 相似文献
87.
88.
Victor S. Sapirstein Charles E. Nolan Itzhak Fischer Elizabeth Cochary Susana Blau Cheryl J. Flynn 《Neurochemical research》1991,16(2):123-128
Plasmolipin is a plasma membrane proteolipid is a major myelin membrane component (Cochary et al., 1990). In this study we report the phylogenic expression of plasmolipin in the vertebrate nervous system. Using Western blot analysis with polyclonal antibodies, we have analyzed membrane fractions, including myelin, from elasmobranchs, teleosts, amphibians, reptiles, birds and mammals. On the basis of immune detection, plasmolipin appears to be restricted to the mammalian nervous system. Comparison of the central and peripheral nervous systems of mammals showed only minor differences in the level of plasmolipin in these two regions. Within mammals, little quantitative differences were observed when rat, human and bovine membrane fractions were compared. The late evolutionary expression of plasmolipin which results in its restriction to mammals makes it unique among the (major) myelin proteins. The potential physiologic significance of these data are discussed.Abbreviations EDTA
Ethylene diamine N.,NN tetracetic acid
- EGTA
Ethylene glycol bis-(B-Aminoethyl Ether) N,,NN tetracetic acid
- MES
([N-Morpholino] ethane sulfonic acid) DCCD, N, Dicyclohexyl carbodiimide 相似文献
89.
David B. Sattelle Sarah C. R. Lummis James F. H. Wong James J. Rauh 《Neurochemical research》1991,16(3):363-374
A GABA-operated Cl– channel that is bicuculline-insensitive is abundant in the nervous tissue of cockroach, in housefly head preparations and thorax/abdomen preparations, and in similar preparations from several insect species. Bicuculline-insensitive GABA-operated Cl– channels, which are rare in vertebrates, possess sites of action of benzodiazepines, steroids and insecticides that are pharmacologically-distinct from corresponding sites on vertebrate GABAA receptors. The pharmacological profile of the benzodiazepine-binding site linked to an insect CNS GABA-operated Cl– channel resembles more closely that of vertebrate peripheral benzodiazepine-binding sites. Six pregnane steroids and certain polychlorocycloalkane insecticides, which are active att-butylbicy-clophosphorothionate (TBPS)-binding sites, also differ in their effectiveness on vertebrate and insect GABA receptors. Radioligand binding and physiological studies indicate that in insects there may be subtypes of the GABA receptor. Molecular biology offers experimental approaches to understanding the basis of this diversity.Special issue dedicated to Dr. Eugene Roberts 相似文献
90.
Pyruvate decarboxylase (PDC) contains thiamine pyrophosphate (TPP) and Mg2+ as cofactors. 31P NMR studies with PDC in the presence of added Mn2+ reveal the pyrophosphate moiety of TPP to be a nonaccessible area for the external Mn2+ and thus proving the Mg-P-complex (taking part in the binding of the coenzyme to the protein) to be a nonaccessible area for the medium. Glyoxylic acid, acting as an inhibitor of PDC by forming a noncleavable bond with the catalytic center of TPP causes a steric immobilization of the coenzyme indicated by a line broadening of the pyrophosphate moiety. 相似文献