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1.
Anti-thrombin activities of heparin. Effect of saccharide chain length on thrombin inhibition by heparin cofactor II and by antithrombin. 总被引:1,自引:0,他引:1
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The interactions of two proteinase inhibitors, heparin cofactor II and antithrombin, with thrombin are potentiated by heparin. Using two methods, we have studied the potentiating effects of a series of heparin (poly)saccharides with high affinity for antithrombin and mean Mr ranging from approx. 1700 to 18,800. First, catalytic amounts of heparin (poly)saccharide were added to purified systems containing thrombin and either heparin cofactor II or antithrombin. Residual thrombin activity was determined with a chromogenic substrate. It was found that only the higher-Mr polysaccharides (Mr greater than 8000) efficiently catalysed thrombin inhibition by heparin cofactor II, there being a progressive catalytic effect with increasing Mr of the polysaccharide. Weak accelerating effects were noted with low-Mr saccharides (Mr less than 8000). This contrasted with the well-characterized interaction of heparin with antithrombin and thrombin, where heparin oligosaccharides of Mr less than 5400 had absolutely no ability to accelerate the reaction, while (poly)saccharides of Mr exceeding 5400 showed rapidly increasing catalytic activity with increasing Mr. Secondly, these and other heparin preparations were added in a wide concentration range to plasma with which 125I-labelled thrombin was then incubated for 30 s. Inhibited thrombin was determined from the distribution of labelled thrombin amongst inhibitor-thrombin complexes, predominantly antithrombin-thrombin and heparin cofactor II-thrombin complexes. In this situation, where the inhibitors competed for thrombin and for the (poly)saccharides, it was found that, provided the latter were of high affinity for antithrombin and exceeded a Mr of 5400, thrombin inhibition in plasma was mediated largely through antithrombin. Polysaccharides of Mr exceeding 8000 that were of low affinity for antithrombin accelerated thrombin inhibition in plasma through their interaction with heparin cofactor II. High concentrations of saccharides of Mr 1700-5400 exhibited a size-dependent acceleration of thrombin inhibition, not through their interaction with antithrombin, but through their interaction with heparin cofactor II. 相似文献
2.
On choice of substrate and habitat in brachionid rotifers 总被引:9,自引:9,他引:0
Information on the distribution of 28 rotifers of the family Brachionidae from diverse waters in south and central Sweden was analyzed to reveal their relationships to substrate and habitat. Some brachionids are preferably planktic, others periphytic and/or benthic. Some non-planktic habitats are utilized more than others, but there is no evidence of a chemical attraction from any substrate. Instead, some substrates seem to be avoided, possibly depending on a poorer flora of periphytic algae. Besides substrate type, the following factors are found to be important for creating separate ecological niches in the brachionid family: temperature, oxygen content, trophic degree, chemical environment, food choice and sensitivity to predation. It is possible to delineate separate ecological niches for all brachionid rotifers, implying that Hutchinson's ideas about the plankton paradox are contradicted. Some species are specialists, other are generalists, the latter being characterized by a great morphological variation. The species are adapted in different ways to their preferential habitats, as regards foot, egg-carrying, protrusions and other lorical structures etc. Longer spines, for instance, are generally found in more transparent water, being a supposed protection against visual predators. 相似文献
3.
Monoclonal antibodies specific for oligosaccharides prepared by partial nitrous acid deamination of heparin 总被引:5,自引:0,他引:5
G Pejler U Lindahl O Larm E Scholander E Sandgren A Lundblad 《The Journal of biological chemistry》1988,263(11):5197-5201
Monoclonal antibodies were raised against a conjugate between heparin oligosaccharides and human serum albumin. The oligosaccharides were prepared by partial nitrous acid degradation of heparin and were coupled to human serum albumin by reductive amination. Characterization of the antibodies secreted by one of the resulting clones showed that they recognize a determinant present in the oligosaccharide antigen, but not in intact heparin, nor in a variety of related polysaccharides. Degradation of heparin by nitrous acid generates a 2,5-anhydro-D-mannose residue at the reducing end of the resulting oligosaccharides, and it is concluded that this structure is essential for interaction with the antibodies. Reduced oligosaccharides (containing a terminal anhydromannitol residue) are also active. After gel chromatography of partially degraded heparin, the smallest components capable of binding to the antibodies were found in a tetrasaccharide fraction. Affinity chromatography on immobilized monoclonal antibodies separated this tetrasaccharide fraction into distinct populations of binding and nonbinding species. Structural analysis showed that the tetrasaccharide fraction that bound to the monoclonal antibodies contained one single component with the structure IdoA(2-OSO3)-GlcNSO3 (6-OSO3)-IdoA(2-OSO3)-aManR(6-OSO3), whereas the fraction that did not bind to the antibodies contained a mixture of different structures. 相似文献
4.
On the ecology of Colurellidae (Rotifera) 总被引:1,自引:1,他引:0
A material consisting of 45 colurellid rotifers from diverse waters in south and central Sweden was analyzed to reveal their relationships to their substrate. Periphytic substrates were generally preferred. Most species are obviously mobile and more or less euryecious, but a few show a predilection for bog environments. Some colurellids are euryhaline, but when inhabiting fresh water they prefer rivers and other environments with a high oxygen concentration, probably because of problems with osmoregulation. 相似文献
5.
Information on the distributions of 77 bdelloid rotifers from diverse waters in south and central Sweden was analyzed to reveal their relationships to substrate and habitat. Only two species were reported from plankton, both in low frequency. An artificial substrate, white cotton, was utilized only by few bdelloids, mainly by species with eyes. Only one species occurred at higher frequency on the saprobic substrate Sphaerotilus. Most bdelloids prefer environments rich in oxygen, but some species may be found in soft-bottom sediments. Several species dwell preferentially in diverse habitats of bogs. Some bdelloids have an exceptionally broad ecological range, but even for these is it possible to distinguish a pattern of preference and avoidance. 相似文献
6.
On the ecology of Dicranophoridae (Rotifera) 总被引:1,自引:1,他引:0
Information on the distribution of 59 dicranophorid rotifers from diverse habitats in south and central Sweden was analyzed to reveal their relationships to their substrate. They were mainly frequenting macrophytic vegetation. Even some species which have previously been regarded as psammobionts were mostly or only found on periphytic substrates. Like other raptorial predators, dicranophorids have a very broad ecological range. 相似文献
7.
Relation to habitat in rotifers 总被引:4,自引:2,他引:2
Rotifera should be especially suited for an analysis of habitat relations because this group contains such a high number of species, inhabiting diverse environments. Furthermore, rotifers are to a large extent cosmopolitan, implying that ecological barriers, rather than geographical, are decisive of their distribution. In this review a short characterization of the rotifer fauna in different habitats is given, whereby macroenvironments and microenvironments are reported separately. The macroenvironments are classified as follows: harmonious lakes and ponds, arctic and antarctic waters, hot springs, hypertrophic-saprobic environments, mires, strongly acidic waters, saline waters, temporary water bodies, subterranean waters, running waters, oceans, terrestrial environments. The following microenvironments are distinguished: macrophytes (housing periphytic rotifers), open water (with planktic forms), minerogenous sediments (with psammon and hyporheos), organogenous sediments, other organisms (i.e. parasites and epizoans).Many rotifers are more or less euryecious, while relatively few are strongly restricted in their choice of habitat. In extreme environments a low number of species is found, but often a high number of individuals within these species. These rotifers are usually primary consumers, and for natural reasons extreme environments are characterized by a low number of trophic levels.In environments with a high species number the separate species differ very much in their morphology, making it difficult to find common traits which may be interpreted as adaptations to the respective habitats. The most apparent adaptations ought to be found among the planktic rotifers, and these adaptations seem to constitute largely a protection against predators. Rotifers in extreme environments are usually not very apart in a morphological or taxonomical respect, with their most close relatives living in normal habitats and sometimes euryecious (an apparent exception from this rule is formed by the class Seisonidea). Adaptations to deviating chemical and physical environments may develop relatively rapidly (seen from a geological perspective), while the more fundamental changes (occurring during a longer period of time) seem to be a response to biotic factors (e.g., the development of different types of trophi for facilitating food collection). 相似文献
8.
On the ecology of Trichocercidae (Rotifera) 总被引:2,自引:2,他引:0
A material consisting of 46 trichocercid rotifers from diverse waters in south and central Sweden was analyzed to reveal their relationships to their substrate. Most species were found in a wide variety of periphytic environments, but a few seemed to be specialized on bogs or psammon, respectively. Several trichocercids are generally regarded as plankters, but some of them obviously invade the open water only on certain occasions. A few species of Trichocerca are registered as indicators of eutrophy, but in some cases only a planktic occurrence may be regarded as indicating. On the other hand, hypertrophic environments are generally avoided. Trichocerca similis prefers polyhumic environments, T. uncinata oligotrophic and running-water conditions. The occurrence is often best explained by studying the food choice. 相似文献
9.
Mast cell tryptase is stored as an active tetramer in complex with heparin in mast cell secretory granules. Previously, we demonstrated the dependence on heparin for the activation/tetramer formation of a recombinant tryptase. Here we have investigated the structural requirements for this activation process. The ability of heparin-related saccharides to activate a recombinant murine tryptase, mouse mast cell protease-6 (mMCP-6), was strongly dependent on anionic charge density and size. The dose-response curve for heparin-induced mMCP-6 activation displayed a bell-shaped appearance, indicating that heparin acts by binding to more than one tryptase monomer simultaneously. The minimal heparin oligosaccharide required for binding to mMCP-6 was 8-10 saccharide units. Gel filtration analyses showed that such short oligosaccharides were unable to generate tryptase tetramers, but instead gave rise to active mMCP-6 monomers. The active monomers were inhibited by bovine pancreatic trypsin inhibitor, whereas the tetramers were resistant. Furthermore, monomeric (but not tetrameric) mMCP-6 degraded fibronectin. Our results suggest a model for tryptase tetramer formation that involves bridging of tryptase monomers by heparin or other highly sulfated polysaccharides of sufficient chain length. Moreover, our results raise the possibility that some of the reported activities of tryptase may be related to active tryptase monomers that may be formed according to the mechanism described here. 相似文献
10.
Henningsson F Ledin J Lunderius C Wilén M Hellman L Pejler G 《Biological chemistry》2002,383(5):793-801
Heparin-deficient mice, generated by gene targeting of N-deacetylase/N-sulfotransferase-2 (NDST-2), display severe mast cell defects, including an absence of stored mast cell proteases. However, the mechanism behind these observations is not clear. Here we show that NDST-2+/+ bone marrow-derived mast cells cultured in the presence of IL-3 synthesise, in addition to highly sulphated chondroitin sulphate (CS), small amounts of equally highly sulphated heparin-like polysaccharide. The corresponding NDST-2-/- cells produced highly sulphated CS only. Carboxypeptidase A (CPA) activity was detected in NDST+/+ cells but was almost absent in the NDST-/- cells, whereas tryptase (mouse mast cell protease 6; mMCP-6) activity and antigen was detected in both cell types. Antigen for the chymase mMCP-5 was detected in NDST-2+/+ cells but not in the heparin-deficient cells. Northern blot analysis revealed mRNA expression of CPA, mMCP-5 and mMCP-6 in both wild-type and NDST-2-/- cells. A approximately 36 kDa CPA band, corresponding to proteolytically processed active CPA, as well as a approximately 50 kDa pro-CPA band was present in NDST-2+/+ cells. The NDST-2-/- mast cells contained similar levels of pro-CPA as the wild-type mast cells, but the approximately 36 kDa band was totally absent. This indicates that the processing of pro-CPA to its active form may require the presence of heparin and provides the first insight into a mechanism by which the absence of heparin may cause disturbed secretory granule organisation in mast cells. 相似文献