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排序方式: 共有107条查询结果,搜索用时 93 毫秒
71.
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73.
Nan Li Peter Yun Mangala A. Nadkarni Nazila Babapoor Ghadikolaee Ky‐Anh Nguyen Mihwa Lee Neil Hunter Charles A. Collyer 《Molecular microbiology》2010,76(4):861-873
Porphyromonas gingivalis is an obligately anaerobic bacterium recognized as an aetiological agent of adult periodontitis. P. gingivalis produces cysteine proteinases, the gingipains. The crystal structure of a domain within the haemagglutinin region of the lysine gingipain (Kgp) is reported here. The domain was named K2 as it is the second of three homologous structural modules in Kgp. The K2 domain structure is a ‘jelly‐roll’ fold with two anti‐parallel β‐sheets. This fold topology is shared with adhesive domains from functionally diverse receptors such as MAM domains, ephrin receptor ligand binding domains and a number of carbohydrate binding modules. Possible functions of K2 were investigated. K2 induced haemolysis of erythrocytes in a dose‐dependent manner that was augmented by the blocking of anion transport. Further, cysteine‐activated arginine gingipain RgpB, which degrades glycophorin A, sensitized erythrocytes to the haemolytic effect of K2. Cleaved K2, similar to that found in extracted Kgp, lacks the haemolytic activity indicating that autolysis of Kgp may be a staged process which is artificially enhanced by extraction of the protein. The data indicate a functional role for K2 in the integrated capacity conferred by Kgp to enable the porphyrin auxotroph P. gingivalis to capture essential haem from erythrocytes. 相似文献
74.
Corn samples were collected in 1999 from three departments of Entre Réos province, Argentina, and were surveyed for mould
contamination and natural occurrence ofFusarium mycotoxins, ochratoxin A and aflatoxins.Fusarium verticillioides was the most prevalent fungal species recorded at all departments. Zearalenone, deoxynivalenol and ochratoxin A were not
found in any samples. Only one of the 52 corn samples analysed was contaminated with aflatoxin B1 (17 μg/kg). Fumonisin B1 was found in 58 % of samples (range of positive samples: 47– 3,347 μg/kg), fumonisin B2 in 33.0 % (range of positive samples: 23–537 μg/kg) and fumonisin B3 in 25.0 % (range of positive samples: 24–287 μg/kg) of them. This is the first report on the natural occurrence of mycotoxins
in corn from Entre Ríos province, Argentina. Levels of fumonisins were lower than detected in other Argentinian provinces. 相似文献
75.
Despite the recent introduction of biological response modifiers and potent new small-molecule antirheumatic drugs, the efficacy of methotrexate is nearly unsurpassed in the treatment of inflammatory arthritis. Although methotrexate was first introduced as an antiproliferative agent that inhibits the synthesis of purines and pyrimidines for the therapy of malignancies, it is now clear that many of the anti-inflammatory effects of methotrexate are mediated by adenosine. This nucleoside, acting at one or more of its receptors, is a potent endogenous anti-inflammatory mediator. In confirmation of this mechanism of action, recent studies in both animals and patients suggest that adenosine-receptor antagonists, among which is caffeine, reverse or prevent the anti-inflammatory effects of methotrexate. 相似文献
76.
Mandal A Liyanage MR Zaidi A Johnson CK 《Protein science : a publication of the Protein Society》2008,17(3):555-562
The Ca2+ signaling protein calmodulin (CaM) stimulates Ca2+ pumping in the plasma-membrane Ca2+-ATPase (PMCA) by binding to an autoinhibitory domain, which then dissociates from the catalytic domain of PMCA to allow full activation of the enzyme. We measured single-molecule fluorescence trajectories with polarization modulation to track the conformation of the autoinhibitory domain of PMCA pump bound to fluorescently labeled CaM. Interchange of the autoinhibitory domain between associated and dissociated conformations was detected at a physiological Ca2+ concentration of 0.15 microM, where the enzyme is only partially active, but not at 25 microM, where the enzyme is fully activated. In previous work we showed that the conformation of the autoinhibitory domain in PMCA-CaM complexes could be monitored by the extent of modulation of single-molecule fluorescence generated with rotating excitation polarization. In the present work, we determined the timescale of association and dissociation of the autoinhibitory domain with the catalytic regions of the PMCA. Association of the autoinhibitory domain was rare at a high Ca2+ concentration (25 microM). At a lower Ca2+ concentration (0.15 microM), conformations of the autoinhibitory domain interchanged with a dissociation rate of 0.042 +/- 0.011 sec(-1) and an association rate of 0.023 +/- 0.006 sec-1. The results indicate that the response time of PMCA upon a reduction in Ca2+ is limited to tens of seconds by autoinhibitory dynamics. This property may reduce the sensitivity of PMCA to transient reductions in intracellular Ca2+. We suggest that the dynamics of the autoinhibitory domain may play a novel role in regulating PMCA activity. 相似文献
77.
Vít Herynek Andrea Gálisová Mangala Srinivas Eric A. W. van Dinther Lucie Kosinová Jiri Ruzicka Markéta Jirátová Jan Kriz Daniel Jirák 《Biological procedures online》2017,19(1):6
Background
In vitro labelling of cells and small cell structures is a necessary step before in vivo monitoring of grafts. We modified and optimised a procedure for pancreatic islet labelling using bimodal positively charged poly(lactic-co-glycolic acid) nanoparticles with encapsulated perfluoro crown ethers and indocyanine green dye via microporation and compared the method with passive endocytosis.Results
Pancreatic islets were microporated using two pulses at various voltages. We tested a standard procedure (poration in the presence of nanoparticles) and a modified protocol (pre-microporation in a buffer only, and subsequent islet incubation with nanoparticles on ice for 10 min).We compared islet labelling by microporation with labelling by endocytosis, i.e. pancreatic islets were incubated for 24 h in a medium with suspended nanoparticles.In order to verify the efficiency of the labelling procedures, we used 19F magnetic resonance imaging, optical fluorescence imaging and confocal microscopy.The experiment confirmed that microporation, albeit fast and effective, is invasive and may cause substantial harm to islets. To achieve sufficient poration and to minimise the reduction of viability, the electric field should be set at 20 kV/m (two pulses, 20 ms each).Poration in the presence of nanoparticles was found to be unsuitable for the nanoparticles used. The water suspension of nanoparticles (which served as a surfactant) was slightly foamy and microbubbles in the suspension were responsible for sparks causing the destruction of islets during poration. However, pre-microporation (poration of islets in a buffer only) followed by 10-min incubation with nanoparticles was safer.Conclusions
For labelling of pancreatic islets using poly(lactic-co-glycolic acid) nanoparticles, the modified microporation procedure with low voltage was found to be safer than the standard microporation procedure. The modified procedure was fast, however, efficiency was lower compared to endocytosis.78.
Mangala D. Manandhar Falak A. Hussaini Randhir S. Kapil Aboo Shoeb 《Phytochemistry》1985,24(1):199-200
A new constituent characterized as 8-acetyl-3,4-dihydroxy-5,7-dimethoxy-2,2-dimethylchroman has been isolated together with alloevodionol-7-methyl ether, 4-methoxy-1-methyl-2(1H)quinolinone, evolitrine, isoevodionol and its methyl ether from the aerial parts of Euodia lunu-ankenda. Its structure was confirmed by its transformation to alloevodionol-7-methyl ether. 4-Methoxy-1-methyl-2(1H)quinolinone and its isomer were synthesized by a modified procedure. 相似文献
79.
The ability of particular cell surface glycoproteins to recycle and become
exposed to individual Golgi enzymes has been demonstrated. This study was
designed to determine whether endocytic trafficking includes significant
reentry into the overall oligosaccharide processing pathway. The Lec1
mutant of Chinese hamster ovary (CHO) cells lack N -
acetylglucosaminyltransferase I (GlcNAc-TI) activity resulting in surface
expression of incompletely processed Man5GlcNAc2 N -linked
oligosaccharides. An oligosaccharide tracer was created by exoglycosylation
of cell surface glycoproteins with purified porcine GlcNAc-TI and
UDP-[3H]GlcNAc. Upon reculturing, all cell surface glycoproteins that
acquired [3H]GlcNAc were acted upon by intracellular mannosidase II, the
next enzyme in the Golgi processing pathway of complex N -linked
oligosaccharides (t1/2= 3-4 h). That all radiolabeled cell surface
glycoproteins were included in this endocytic pathway indicates a common
intracellular compartment into which endocytosed cell surface glycoproteins
return. Significantly, no evidence was found for continued oligosaccharide
processing consistent with transit through the latter cisternae of the
Golgi apparatus. These data indicate that, although recycling plasma
membrane glycoproteins can be reexposed to individual Golgi-derived
enzymes, significant reentry into the overall contiguous processing pathway
is not evident.
相似文献
80.
The role of site-specific N-glycosylation in secretion of soluble forms of rabies virus glycoprotein 总被引:1,自引:1,他引:0
Wojczyk BS; Stwora-Wojczyk M; Shakin-Eshleman S; Wunner WH; Spitalnik SL 《Glycobiology》1998,8(2):121-130
Rabies virus glycoprotein is important in the biology and pathogenesis of
neurotropic rabies virus infection. This transmembrane glycoprotein is the
only viral protein on the surface of virus particles, is the viral
attachment protein that facilitates virus uptake by the infected cell, and
is the target of the host humoral immune response to infection. The
extracellular domain of this glycoprotein has N- glycosylation sequons at
Asn37, Asn247, and Asn319. Appropriate glycosylation of these sequons is
important in the expression of the glycoprotein. Soluble forms of rabies
virus glycoprotein were constructed by insertion of a stop codon just
external to the transmembrane domain. Using site-directed mutagenesis and
expression in transfected eukaryotic cells, it was possible to compare the
effects of site-specific glycosylation on the cell-surface expression and
secretion of transmembrane and soluble forms, respectively, of the same
glycoprotein. These studies yielded the surprising finding that although
any of the three sequons permitted cell surface expression of full-length
rabies virus glycoprotein, only the N-glycan at Asn319 permitted secretion
of soluble rabies virus glycoprotein. Despite its biological and medical
importance, it has not yet been possible to determine the crystal structure
of the full-length transmembrane form of rabies virus glycoprotein which
contains heterogeneous oligosaccharides. The current studies demonstrate
that a soluble form of rabies virus glycoprotein containing only one sequon
at Asn319 is efficiently secreted in the presence of the N-glycan
processing inhibitor 1-deoxymannojirimycin. Thus, it is possible to purify
a conformationally relevant form of rabies virus glycoprotein that contains
only one N-glycan with a substantial reduction in its microheterogeneity.
This form of the glycoprotein may be particularly useful for future studies
aimed at elucidating the three-dimensional structure of this important
glycoprotein.
相似文献