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81.
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83.
Engineering receptor-mediated cytotoxicity into human ribonucleases by steric blockade of inhibitor interaction 总被引:3,自引:0,他引:3
Suzuki M Saxena SK Boix E Prill RJ Vasandani VM Ladner JE Sung C Youle RJ 《Nature biotechnology》1999,17(3):265-270
Several nonmammalian members of the RNase A superfamily exhibit anticancer activity that appears to correlate with resistance to the cytosolic ribonuclease inhibitor (RI). We mutated two human ribonucleases-pancreatic RNase (hRNAse) and eosinophil-derived neurotoxin (EDN)-to incorporate cysteine residues at putative sites of close contact to RI, but distant from the catalytic sites. Coupling of Cys89 of RNase and Cys87 of EDN to proteins at these sites via a thioether bond produced enzymatically active conjugates that were resistant to RI. To elicit cellular targeting as well as to block RI binding, transferrin was conjugated to a mutant human RNase, rhRNase(Gly89)-->Cys) and a mutant EDN (Thr87-->Cys). The transferrin-rhRNase(Gly89-->Cys) thioether conjugate was 5000-fold more toxic to U251 cells than recombinant wild-type hRNase. In addition, transferrin-targeted EDN exhibited tumor cell toxicities similar to those of hRNase. Thus, we endowed two human RI-sensitive RNases with greater cytotoxicity by increasing their resistance to RI. This strategy has the potential to generate a novel set of recombinant human proteins useful for targeted therapy of cancer. 相似文献
84.
The ultrastructure of prostate gland of Lampito mauritii revealed two types of secretory cells. Type 1 cells with a broad basal region and a long apical region contain electron dense oval secretory granules with an increased density at the core region. Numerous electron lucent granules with fine filamentous and electron dense amorphous materials also occur at the basal region of these cells. Type 2 cells contain electron lucent mucous-like secretory granules. This cell type contains exceptionally large Golgi complexes having 20-23 stacked cisternae. Both cell types open into a common lumen and numerous microtubules are visible at the apical end. Junctional complexes, such as desmosomes and septate junctions, are observed in this glandular tissue. 相似文献
85.
J.M.R. Victor B.N.S. Murthy S.J. Murch S. KrishnaRaj P.K. Saxena 《Plant Growth Regulation》1999,28(1):41-47
Somatic embryogenesis was induced at the hypocotyledonary notch region of intact peanut (Arachis hypogaea L.) seedings cultured on a medium containing 10 mol·L–1 thidiazuron (TDZ). Inclusion of the purine analogs 2,6-diaminopurine (DAP), azaadenine or azaguanine to the thidiazuron amended medium inhibited the embryogenic response of the seedlings. DAP-mediated inhibition was not overcome by the addition of adenine sulphate. Inhibition of the embryogenic response by DAP provides evidence that the TDZ-induced accumulation of purine cytokinins is an essential component of TDZ-induced somatic embryogenesis process. Analyses of the endogenous level of purine metabolites indicated that supplementation of the media with TDZ resulted in an overall increase in the endogenous cytokinins while DAP inhibited the purine recycling resulting in decreased levels of endogenous adenine and zeatin. 相似文献
86.
Sengupta K Banerjee S Saxena NK Jonathan NB Campbell DR Banerjee SK 《Molecular and cellular biochemistry》2004,262(1-2):215-224
Beta-estradiol (17beta-E2) augments VEGF-A expression in various estrogen targeted organs and cells including breast tumor derived cell lines, via an ER-alpha mediated pathway. Ironically, 17beta-E2 is able to regulate some genes via ER-alpha independent pathways. In the present study, we sought to determine whether 17beta-E2 can modulate VEGF-A expression in absence of ER-alpha, and therefore, three different cell lines including ER-alpha+ MCF-7, and ER-alpha SKBR-3 and HMEC were used for this study. The present study demonstrates that 17beta-E2 also induces VEGF-A mRNA expression in ER-negative SKBR-3 breast tumor cells in a manner similar to that observed in ER-positive MCF-7 cells. Blocking the induced-expression by antiestrogen ICI 182,780 indicates the induction pathway is ER dependent. While ER-alpha mRNA is absent in both HMEC and SKBR-3 cells, the impact of estrogen was found only in SKBR-3 cells, suggesting the existence of an analogue to ER-alpha or overlapping signal in these cells. Consistent with this suggestion, the present studies demonstrate the existence of an ER-alpha(var2) protein in MCF-7 and in SKBR-3 cells. This variant is predominantly localized in the nuclei of SKBR-3 cells. Importantly, specific binding of 17beta-E2 by these cells suggest the ER-alpha(var2) may act as active receptor in SKBR-3 cells. 相似文献
87.
Ikegami T Verdier L Sakhaii P Grimme S Pescatore B Saxena K Fiebig KM Griesinger C 《Journal of biomolecular NMR》2004,29(3):339-349
A molecule with an anisotropic magnetic susceptibility is spontaneously aligned in a static magnetic field. Alignment of such a molecule yields residual dipolar couplings and pseudocontact shifts. Lanthanide ions have recently been successfully used to provide an anisotropic magnetic susceptibility in target molecules either by replacing a calcium ion with a lanthanide ion in calcium-binding proteins or by attaching an EDTA derivative to a cysteine residue via a disulfide bond. Here we describe a novel enantiomerically pure EDTA derived tag that aligns stronger due to its shorter linker and does not suffer from stereochemical diversity upon lanthanide complexation. We observed residual (15)N,(1)H-dipolar couplings of up to 8 Hz at 800 MHz induced by a single alignment tensor from this tag. 相似文献
88.
Agnihotri VK Thappa RK Meena B Kapahi BK Saxena RK Qazi GN Agarwal SG 《Phytochemistry》2004,65(16):2411-2413
Fresh aerial parts of Angelica glauca, growing wild in Kashmir valley in higher Himalaya (Jammu and Kashmir, India), collected at flowering stage from different locations, on hydro-distillation provided a refreshing light pale coloured essential oil with characteristic floral woody flavour. The oil was found to be a complex mixture of mono- and sesquiterpenes and 34 compounds accounting for nearly 97.4% of the oil were characterized with the help of capillary GC, GC-MS, and NMR. Major compounds of the oil were characterized as alpha-phellandrene (13.5%), trans-carveol (12.0%), beta-pinene (11.7%), thujene (7.5%), beta-caryophyllene oxide (7.2%), beta-caryophyllene (7.0%), gamma-terpinene (6.7%), nerolidol (6.5%), beta-bisabolene (5.2%) and germacrene D (4.5%). It is the first report to exploit the essential oil from Himalayan A. glauca herb collected at flowering stage. 相似文献
89.
Cyanobacteria are versatile tetrapyrrole synthesizers that can regulate their tetrapyrrole content and composition in response to environmental signals. The present investigation analyses the interplay between light and dark regimes (continuous light, light-dark cycles (16:8) and continuous darkness) and aerobic, air-tight, and anaerobic environments (argon-enriched), on the relative composition of various pigments and growth attributes of Calothrix elenkenii as a prelude to exploiting C. elenkenii's bioindustrial potential as a source of pigments. Incubation in an anaerobic environment stimulated hormogonia formation and induced colouration/thickening of cells. Aerobically grown cultures of Calothrix, under continuous illumination produced the maximum amount of total phycobiliproteins and sugars, although chlorophyll accumulation and nitrogenase activity were highest in the light-dark environment. However, the beta-carotene content was observed to vary under anaerobic conditions with different light-dark regimes. This C. elenkenii strain can be a valuable source of pigments under optimized environmental conditions. 相似文献
90.
Kamal A Ramu R Khanna GB Saxena AK Shanmugavel M Pandita RM 《Bioorganic & medicinal chemistry letters》2004,14(19):4907-4909
New pyrrolobenzodiazepine-anthraquinone hybrids have been designed and synthesized, found to effectively bind to DNA and also exhibit cytotoxicity against many cancer cell lines 相似文献