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91.
92.
A. Cortes D. C. Emery D. J. Halsall R. M. Jackson A. R. Clarke J. J. Holbrook 《Protein science : a publication of the Protein Society》1992,1(7):892-901
The proposal that the active site vacuole of NAD(+)-S-lactate dehydrogenase is unable to accommodate any imbalance in electrostatic charge was tested by genetically manipulating the cDNA coding for human muscle lactate dehydrogenase to make a protein with an aspartic acid introduced at position 140 instead of the wild-type asparagine. The Asn 140-Asp mutant enzyme has the same kcat as the wild type (Asn 140) at low pH (4.5), and at higher pH the Km for pyruvate increases 10-fold for each unit increase in pH up to pH 9. We conclude that the anion of Asp 140 is completely inactive and that it binds pyruvate with a Km that is over 1,000 times that of the Km of the neutral, protonated aspartic-140. Experimental results and molecular modeling studies indicate the pKa of the active site histidine-195 in the enzyme-NADH complex is raised to greater than 10 by the presence of the anion at position 140. Energy minimization and molecular dynamics studies over 36 ps suggest that the anion at position 140 promotes the opening of and the entry of mobile solvent beneath the polypeptide loop (98-110), which normally seals off the internal active site vacuole from external bulk solvent. 相似文献
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94.
95.
Twila Jackson Michael F. Allard Catherine M. Sreenan Lisa K. Doss Sanford P. Bishop Judith L. Swain 《Molecular and cellular biochemistry》1991,104(1-2):15-19
Transgenic animals provide a model system to elucidate the role of specific proteins in development. This model is now being used increasingly in the cardiovascular system to study cardiac growth and differentiation. During cardiac myocyte development a transition occurs from hyperplastic to hypertrophic growth. In the heart the switch from myocyte proliferation to terminal differentiation is synchronous with a decrease in c-myc mRNA abundance. To determine whether c-myc functions to regulate myocyte proliferation and/or differentiation, we examined the in vivo effect of increasing c-myc expression during fetal development and of preventing the decrease in c-myc mRNA expression that normally occurs during myocyte development. The model system used was a strain of transgenic mice exhibiting constitutive expression of c-myc mRNA in cardiac myocytes throughout development. Increased c-myc mRNA expression is associated with both atrial and ventricular enlargement in the transgenic mice. This increase in cardiac mass is secondary to myocyte hyperplasia, with the transgenic hearts containing greater than twice as many myocytes as nontransgenic hearts. The results of this study indicate that constitutive expression of c-myc mRNA in the heart during development results in enhanced hyperplastic growth, and suggest a regulatory role for the c-myc protooncogene in cardiac myogenesis. 相似文献
96.
Cytoskeletal proteins in adherens-type cell-matrix junctions. 总被引:1,自引:0,他引:1
D R Critchley A Gilmore L Hemmings P Jackson A McGregor V Ohanian B Patel G Waites C Wood 《Biochemical Society transactions》1991,19(4):1028-1033
97.
Toshiyuki Itaya Eric Fearon Troy Fiesinger Barbara Hunt Bert Vogelstein Philip Frost 《Cancer immunology, immunotherapy : CII》1991,33(4):267-273
Summary The transfection of murine SP1 tumor cells with the hemagglutinin (HA) gene of influenza virus results, after fluorescent-activated cell sorting (FACS), in the selection of high-HA-expressing cell lines called H4A and H4B. Both lines fail to grow in syngeneic animals at doses that result in 100% tumor take of non-transfected tumor cells. Both grow in immunosuppressed mice. SP1 and H4A or H4B cells express few class I major histocompatibility complex (MHC) antigens but do express class II IAk antigens. H4A or H4B cells engender a cytotoxic T lymphocyte (CTL) response but cannot protect against a challenge with SP1 cells. This CTL response is inhibited by anti-CD4 but not anti-CD8 antibodies. Using FACS, we were able to select a population (called H5AK5) with high class-I MHC antigen expression. Like H4A and H4B, H5AK5 cells fail to grow in syngeneic animals but do grow in immunosuppressed mice. However, unlike H4A or H4B, H5AK5 can induce protection against a challenge with 1 × 105 SP1 cells. These studies indicate that the immunogenicity ofHA-transfected SP1 cells may correlate with the cell-surface expression of class II MHC antigens. However, HA-expressing SP1 cells seem able to induce a protective response against a parent SP1 cell challenge only if they also express class I MHC antigens. This view is supported by the observations that SP1 cells expressing murine interleukin-2 do not express class I MHC antigens, fail to grow in syngeneic animals, do grow in immunosuppressed mice but do not protect against a challenge with parental SP1 cells.This work was supported by The Clayton Fund, The Sid W. Richardson Foundation and PHS grants CA 39853 and 41525. Toshiyuki Itaya is a visiting scientist supported by the Smith Education Fund of the Department of Cell Biology. Troy Fiesinger is a summer research investigator sponsored by The University of Texas M. D. Anderson Cancer Center Summer Program for College Students 相似文献
98.
Philip A. Chiverton 《Entomologia Experimentalis et Applicata》1988,47(2):173-182
Maximum consumption rates were determined for two carabids, Bembidion lampros Herbst. and Pterostichus cupreus L., feeding on the cereal aphid Rhopalosiphum padi L. at different temperatures in the laboratory. Mean daily consumption increased with increasing temperature for both species, B. lampros consuming a maximum of 16 1–3 instar nymphs and 9 apterous adult aphids at 25°C. P. cupreus was particularly voracious and consumed 125 apterous adult R. padi per day at 20°C. The behaviour of both species was analysed by video filming starved beetles, maintained at different constant temperatures, in arenas sown with spring barley. The behavioural components (1) still; (2) run/walk; (3) search and (4) confrontation were identified and were common to both species. P. cupreus was more active over the temperature range tested; B. lampros was inactive under 10°C. The proportion of time spent searching, number of plants searched, and velocity increased with increasing temperature for both species. When observed in similar arenas seeded with R. padi colonies, individuals of P. cupreus significantly increased their time spent searching in arenas with increasing aphid density. Following discovery of an aphid colony, individuals climbed and searched the host plant and its nearest neighbours. Plants in aphid free arenas were rarely climbed. B. lampros was not observed climbing in either aphid free arenas or in arenas with increasing aphid densities, and did not significantly increase its time spent searching in response to increased prey density. The few B. lampros that found aphids caught them walking on the soil surface. The relative efficiences of these two carabids as predators of R. padi are discussed, and the results are compared with similar studies elsewhere with predators of Sitobion avenae on winter wheat.
Résumé Les taux maximum de consommation de R. padi L. à différentes températures ont été déterminés au laboratoire chez deux carabes, B. lampros Herbstet P. cupreus L. La consommation moyenne a augmenté avec la température chez les deux espèces, B. lampros consommant un maximum de 15,9 larves des stades 1 à 3 et 9,1 pucerons adultes aptères, à 25°C. P. cupreus a été particulièrement vorace et a consommé 125,3 adultes aptères par jour à 20°C. Le comportement des deux espèces a été observé en filmant en vidéo des carabes à jeun, maintenus à différentes températures constantes, dans des enceintes semées en orge de printemps. Des éléments du comportement, communs aux deux espèces, ont été définis: 1) immabilité, 2) marche et course, 3) recherche, 4) affrontement. P. cupreus a été plus actif à toutes les températures, B. lamprosa été inactif au-dessous de 10°C. La part de temps consacrée à la recherche, le nombre de plantes prospectées, et la vitesse ont augmenté avec la température chez les deux espèces. Dans des enceintes similaires colonisées par R. padi, P. cupreus a significativement augmenté le temps consacré à la recherche dans les enceintes, parallèlement à l'augmentation de la densité des pucerons. Après la découverte d'une colonie de pucerons, P. cupreus escalade et prospecte la plante et ses voisines immédiates; tandis que les plantes des enceintes sans pucerons sont rarement escaladées. B. lampros n'a pas été observé escaladant des plantes d'enceintes avec ou sans pucerons, et il n'a pas accru son temps de prospection en fonction de la densité de pucerons. Les quelques B. lampros qui ont capturé des pucerons l'ont fait lorsque ceux-ci marchaient sur la surface du sol. La discussion a porté sur l'efficacité relative des deux carabes comme prédateurs de R. padi, et les résultats ont été comparés à ceux d'études du même type, menées ailleurs, avec des prédateurs de Sitobion avenae sur blé d'hiver.相似文献
99.
We studied the effects of 17 beta-estradiol (E2) on luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release induced by drugs that activate different intracellular signal transduction mechanisms in rat anterior pituitary cells. Cells were pretreated with E2 (6 x 10(-10) M) or diluent for 24 h. Then, both E2- and diluent-pretreated cells were incubated for 4 h with E2 or diluent, respectively, with or without drugs, and in the presence or absence of gonadotropin-releasing hormone (GnRH). Media were assayed for LH and FSH by radioimmunoassays. E2 treatment had no effect on basal FSH release, but occasionally stimulated basal LH release. Phospholipase C (PLC), L-alpha-1,2-dioctanoyl glycerol (C8), veratridine, 8-bromo-cyclic adenosine 3',5'-monophosphate (8-Br-cAMP), melittin (a phospholipase A2 [PLA2] activator), arachidonic acid, PLA2, and GnRH all stimulated LH and FSH release in both E2- and diluent-treated cells. E2 treatment increased both LH and FSH release induced by GnRH, PLC, C8, veratridine, and 8-Br-cAMP, but not by melittin, arachidonic acid, and PLA2. Neither C8, PLA2, nor arachidonic acid in combination with a maximal dose of GnRH had additive effects on either LH or FSH release, whereas melittin increased the maximal response to GnRH in both E2- and diluent-treated cells. The effects of veratridine and 8-Br-cAMP depended on dose of GnRH and presence or absence of E2. These results suggest that E2 augments stimulus-coupled gonadotropin release by interacting with the Ca2+-, and/or diacylglycerol-, and cAMP-activated pathways, but not with the arachidonic acid-activated pathway. 相似文献
100.
Philip A. Dennis 《Economic botany》1988,42(1):16-28
Medicinal plants identified by Miskito informants in Awastara, Nicaragua, were collected in the field. They are listed and botanically identified in this paper. Particularly interesting among the collection of 23 plant species are those used to cure snakebite and athlete’s foot, as observed in the field. 相似文献