The transmembrane protein of human immunodeficiency virus type 1 (HIV-1) contains a leucine zipper-like (hydrophobic heptad) repeat which has been predicted to form an amphipathic alpha helix. To evaluate the potential of the hydrophobic heptad repeat to induce protein oligomerization, this region of gp41 has been cloned into the bacterial expression vector pRIT2T. The resulting plasmid, pRIT3, expresses a fusion protein consisting of the Fc binding domain of monomeric protein A, a bacterial protein, and amino acids 538 to 593 of HIV-1 gp41. Gel filtration chromatography demonstrated the presence of oligomeric forms of the fusion protein, and analytical centrifugation studies confirmed that the chimeric protein formed a higher-order multimer that was greater than a dimer. Thus, we have identified a region of HIV-1 gp41 which is capable of directing the oligomerization of a fusion protein containing monomeric protein A. Point mutations, previously shown to inhibit the biological activity of the HIV-1 envelope glycoprotein, have been engineered into the segment of gp41 contained in the fusion protein, and expressed mutant proteins were purified and analyzed via fast protein liquid chromatography. A point mutation in the heptad repeat, which changed the central isoleucine to an alanine, caused a significant (> 60%) decrease in oligomerization, whereas changing the central isoleucine to aspartate or proline resulted in almost a complete loss of oligomerization. Deletions of one, two, or three amino acids following the first isoleucine also resulted in a profound decrease in oligomerization. The inhibitory effects of the mutations on oligomer formation correlated with the effects of the same mutations on envelope glycoprotein-mediated fusion. A possible role of the leucine zipper-like region in the fusion process and in an oligomerization event distinct from assembly of the envelope glycoprotein complex is discussed. 相似文献
Cyclopiazonic acid was shown to be mutagenic to Salmonella typhimurium TA98 and TA100 in the presence of metabolic activation. The activity of cyclopiazonic acid in the presence of aflatoxin B1 was studied in complete factorial experiments with strain TA98. Both mycotoxins produced significant mutagenic activity and in combination. The activity in combination appeared to be additive rather than synergistic. The specific activity of cyclopiazonic acid was estimated to be approximately 140 revertants per mu mol in strain TA98. 相似文献
Treatment of carrot roots (Daucus carota L.) with 10 microliters per liter ethylene in O2 evokes a three- to four-fold increase in polysome prevalence and associated poly(A)+ RNA. The increase in polysome prevalence is attended by a similar change in CO2 evolution. The increase in polysomal poly(A)+ mRNA constitutes primarily a generic increase in constitutive mRNAs as assayed by in vitro translation. However, changes in the relative abundance of several in vitro translatable ethylene specific mRNAs do occur.
Cyanide, at concentrations which inhibit cytochrome oxidase, initiates a respiratory rise very similar in kinetics and magnitude to that evoked by ethylene. Moreover, the combined treatment with cyanide and ethylene evokes a respiratory response resembling that caused by ethylene or cyanide alone. Nevertheless, cyanide, in the presence of ethylene, significantly inhibits the increase in polysome prevalence and new gene expression associated with ethylene treatment of carrot roots. Separation of in vitro translation products by one-dimensional and two-dimensional gel electrophoresis shows that several new in vitro translation products appear in cyanide-treated carrots different from those evoked by ethylene. Engagement of the less energy efficient alternative electron transport path by cyanide may be responsible for inhibition of the normal ethylene associated increase in polysome prevalence and new gene expression. The implications of these results on regulation of respiratory metabolism are discussed and compared with the results for similar experiments with avocado fruit (Tucker and Laties 1984 Plant Physiol 74: 307-315) in which cyanide does not inhibit an ethylene educed increase in polysome prevalence and change in gene expression.
Ovulation was successfully induced with luteinising hormone releasing hormone in 28 women with hypothalamic amenorrhoea who had failed to respond to treatment with clomiphene. Luteinising hormone releasing hormone was administered in a pulsatile manner with miniaturised automatic infusion systems. The rate of ovarian follicular maturation, as monitored by serial pelvic ultrasonography, was similar to that observed in spontaneous cycles. Endocrine assessment by serial measurement of gonadotrophin, oestradiol, and progesterone concentrations showed hormone concentrations to be within the normal range. Intravenous treatment was required in only two patients, the remainder responding satisfactorily to subcutaneous infusion. All patients conceived within six cycles of treatment, and only one multiple pregnancy occurred. 相似文献
Vitamin D intakes of infants aged 6 and 18 months from the Asian community in Southhall, Middlesex, were studied to assess the effectiveness of food fortification as a means of preventing vitamin D deficiency. Infants aged 6 months generally had similar diets to white children of the same age and had reasonable vitamin D intakes owing to consumption of fortified dried milks and cereals, reinforced by health visitors and baby clinics. Children aged 18 months, however, ate largely Asian diets and had much lower vitamin D intakes than the 6-month-old group with a corresponding increase in symptoms of vitamin D deficiency. Hence new measures for preventing vitamin D deficiency should probably be aimed at children aged over 1 year. The results of this survey suggest that fortifying chapati flour would be the most effective method of doing this. 相似文献
A difference Fourier synthesis of deoxyhaemoglobin Tacoma minus deoxyhaemoglobin A at 3.5 Å resolution has been calculated. The map shows a large negative peak due to the removal of the guanidinium group of Arg B12(30)β, surrounded by positive and negative peaks indicative of some atoms moving towards, and others away from, the vacated site. Among the latter, the most important is the carboxylate of Glu B8(26)β which is hydrogen-bonded to the guanidinium group of the arginine in haemoglobin A, but swings round its α-β carbon bond towards the imidazoles of histidines G18(116) and 19(117)β in haemoglobin Tacoma. This movement would raise the pK values of the histidines, so that their positive charges compensated for the loss of the arginine. This may explain why haemoglobin Tacoma has the same electrophoretic mobility as haemoglobin A. It is shown that haemoglobin Tacoma has a lower oxygen equilibrium constant KT and a larger allosteric constant L than haemoglobin A. The lowering of KT may be due to the loosening of the T structure in haemoglobin Tacoma consequent upon the removal of the hydrogen bonds made by the guanidinium group of Arg B12(30)β at the α1β1 contact. Their removal also accounts for the decreased stability of haemoglobin Tacoma. We cannot yet explain its diminished Bohr effect, nor the increase in L. 相似文献
A consideration of all the available data on Triassic vertebrate faunas, and their stratigraphic location reveals a relatively sudden extinction event among the last of the mammal-like reptiles and the herbivorous rhynchosaurs in the Norian of the Upper Triassic. This event was apparently quickly followed by the radiation of the dinosaurs, also in the Norian. This conclusion suggests that competition was not the main factor in the initial success of the dinosaurs, but opportunistic radiation following the extinction of major reptile groups. A global review of Triassic sedimentary facies shows that there were climatic and floral changes towards the end of the Triassic. It is envisaged that increasing aridity in the later Triassic, resulting from plate motions and particularly affecting Gondwanaland and southwestern Laurasia, brought about floral changes and then the reptile extinctions. With the rapid evolution of new floras of conifers and bennettitaleans, the dinosaurs came to dominate all terrestrial faunas within the space of only a few million years. 相似文献
Myocardial revascularization has been carried out by us in 67 patients 70 years of age or older. Advanced coronary artery disease was found at angiography in more than two thirds of the patients. The postoperative morbidity and mortality compare very favorably with those in younger patients. The early and late mortality in the 67 patients was 4.5 percent and 6.0 percent, respectively. Fifty-seven survivors have been followed an average of 21 months; for most patients there has been a pronounced improvement in clinical classification. Properly selected, patients of advanced age can undergo successful revascularization surgical procedures. The adequacy of function of the left ventricle, proper timing of the surgical operation and an aggressive yet realistic approach seem to be major determinants for a good result. 相似文献