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1.
Demonstration of structural differences between the two subunits of human-plasma fibronectin in the carboxy-terminal heparin-binding domain 总被引:6,自引:0,他引:6
H Pande J Calaycay T D Lee K Legesse J E Shively A Siri L Borsi L Zardi 《European journal of biochemistry》1987,162(2):403-411
Structural differences between the two subunits of human plasma fibronectin were studied by analyzing the carboxy-terminal heparin-binding domain (Hep-2). Two fragments (29 kDa and 38 kDa) derived from the Hep-2 domain were purified from thermolysin-digested human plasma fibronectin. Identical NH2-terminal sequences were obtained for both fragments through 16 Edman cycles. Neither domain contained the 90-amino-acid extra domain which is predicted by cDNA analysis of the cellular form of fibronectin. We have examined the primary structures of the 29-kDa and 38-kDa Hep-2 domains produced from the two chains of plasma fibronectin by analyzing the tryptic peptides by fast atom bombardment/mass spectrometry and comparison with the predicted fragments deduced from the corresponding cDNA-derived peptide sequences. Peptides that were unique to each domain were further characterized by microsequence analysis. The two domains showed identical amino acid sequences through 274 residues, followed by a region of variability. The 29-kDa domain contains 279 amino acids with an estimated relative molecular mass (Mr) of 30,460. This domain is located in the heavy chain of plasma fibronectin and contains three repeats of type III sequences plus a portion of the connecting segment (IIICS) region. The 38-kDa domain contains 359 amino acids and one O-linked glycosyl unit for an estimated Mr of 39,263. This domain is from the light chain of plasma fibronectin and contains four repeats of type III sequences with the deletion of the entire 120-amino-acid IIICS area. Secondary structure analysis by Chou/Fasman and circular dichroism reveals extensive beta-sheet structure for these domains. Key sulfhydryl and glycosylation sites are located near the mRNA splice junctions for the two chains. It is postulated that the splice junctions are adjacent to a flexible domain joining two regions of extensive beta-sheet structure. 相似文献
2.
The domain structure of human fibronectins isolated from plasma and from the conditioned medium of normal and transformed fibroblasts was analyzed by limited proteolysis and S-cyanylation followed by immunostaining of released fragments with five kinds of antibodies, each specific for one functional domain. The results indicate that all three human fibronectins are composed of the same set of functional domains aligned in the same topological order. However, the following clear differences were found in specific fragments released from plasma fibronectin (pFN) and those released from fibronectin of normal (N-cFN) and transformed fibroblasts (T-cFN). Two fragments (Mr = 70,000 and 60,000) were released from the COOH-terminal region of pFN by cathepsin D. These fragments represent the COOH-terminal heparin-binding (Hep-2) and fibrin-binding (Fib-2) domains. The corresponding fragments released from both N-cFN and T-cFN by cathepsin D had much larger molecular weights (Mr = 100,000 and 83,000-74,000) than those from pFN. The fragments from the Fib-2 domain alone, however, did not show any difference among all three FNs. The internal region, from the gelatin-binding (Gel) domain through the Hep-2 domain, of N-cFN and T-cFN was released as a Mr = 210,000 fragment upon mild trypsin digestion. The corresponding fragment from pFN was released as a Mr = 185,000 fragment. The COOH-terminal half, including the Hep-2 domain, of both N-cFN and T-cFN was released by S-cyanylation as Mr = 160,000-145,000 fragments, which are 25,000-20,000 larger than the corresponding fragments of pFN. These results clearly indicate that the Hep-2 domain of N-cFN and T-cFN is 30,000-20,000 daltons larger than the same domain of pFN. Although various fragments released from N-cFN and T-cFN showed a similar pattern, there were minor differences. Thermolysin fragments derived from the Hep-2 domain of N-cFN were clearly distinguishable from those from T-cFN. Three groups of fragments with Mr = 40,000, 35,000-32,000, and 30,000 were released from N-cFN, while only the 35,000-32,000 fragment was released from T-cFN. The Mr = 44,000/60,000 thermolysin fragments representing the Gel domain and the Mr = 210,000/165,000 tryptic fragments representing the internal domains of T-cFN were slightly, but consistently, larger than those of N-cFN.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
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4.
Verónica Alvarez Maubecin Viviana N. Sanchez Marcelo D. Rosato Siri Bruce D. Cherksey Mutzuyuki Sugimori Rodolfo Llinás Osvaldo D. Uchitel 《Journal of neurochemistry》1995,64(6):2544-2551
Abstract: The voltage-dependent calcium channels present in mammalian and chicken brain synaptosomes were characterized pharmacologically using specific blockers of L-type channels (1,4-dihydropyridines), N-type channels (ω-conotoxin GVIA), and P-type channels [funnel web toxin (FTX) and ω-agatoxin IVA]. K+-induced Ca2+ uptake by chicken synaptosomes was blocked by ω-conotoxin GVIA (IC50 = 250 nM). This toxin at 5 µM did not block Ca2+ entry into rat frontal cortex synaptosomes. FTX and ω-agatoxin IVA blocked Ca2+ uptake by rat synaptosomes (IC50 = 0.17 µl/ml and 40 nM, respectively). Likewise, in chicken synaptosomes, FTX and ω-agatoxin IVA affected Ca2+ uptake. FTX (3 µl/ml) exerted a maximal inhibition of 40% with an IC50 similar to the one obtained in rat preparations, whereas with ω-agatoxin IVA saturation was not reached even at 5 µM. In chicken preparations, the combined effect of saturating concentrations of FTX (1 µl/ml) and different concentrations of ω-conotoxin GVIA showed no additive effects. However, the effect of saturating concentrations of FTX and ω-conotoxin GVIA was never greater than the one observed with ω-conotoxin GVIA. We also found that 60% of the Ca2+ uptake by rat and chicken synaptosomes was inhibited by ω-conotoxin MVIID (1 µM), a toxin that has a high index of discrimination against N-type channels. Conversely, nitrendipine (10 µM) had no significant effect on Ca2+ uptake in either the rat or the chicken. In conclusion, Ca2+ uptake by rat synaptosomes is potently inhibited by different P-type Ca2+ channel blockers, thus indicating that P-type channels are predominant in this preparation. In contrast, Ca2+ uptake by chicken synaptosomes is sensitive to ω-conotoxin GVIA, FTX, ω-agatoxin IVA, and ω-conotoxin MVIID. This suggests that a channel subtype with a mixed pharmacology is present in chicken synaptosomes. 相似文献
5.
Lynn Siri Kimsey 《Animal behaviour》1980,28(4):996-1004
The male territories of two species of orchid bees, Eulaema meriana and Euglossa imperialis, are described. These territories consist of a perch, where the males display on the trunk of a tree, and a route flown from and back to the perch. Territories are located in treefalls or other large light gaps in tropical forest. The territories of these two species differ in the height of the perch and amount of light reaching it, size of the perch tree, and period of activity. A definition of lek is given. These male euglossines form facultative leks in large treefalls. The relationships between male territories, female foraging ranges, and collection of aromatic material by males are discussed. 相似文献
6.
Fibronectin: a chromatin-associated protein? 总被引:14,自引:0,他引:14
We have previously reported that chromatin preparations from human cultured fibroblasts contain a single homologous serum protein. In this paper we present evidence, based on immunological identity and physicochemical properties, that this serum protein is fibronectin. Furthermore, using a radioimmunoassay system, we have estimated that fibronectin represents about 0.7% of the total protein in both chromatin preparations and whole fibroblasts. Using a nitrocellulose filter assay system, we also show that fibronectin is a DNA-binding protein having an equilibrium constant of 4.6 x 10(-6) M. Equilibrium competition experiments have demonstrated that fibronectin has the ability to differentiate among nucleotides, indicating that fibronectin-DNA interaction is at least partially specific, and that a minimum polymer length of 12-18 nucleotides is required for effective binding to occur. Fibronectin has been isolated readily from plasma using DNA-affinity chromatography. We do not have direct evidence that fibronectin is an actual nonhistone chromosomal protein, but fibronectin is a DNA-binding protein (at least under in vitro assay conditions) and appears to be a normal constituent of chromatin as chromatin is currently isolated from cell nuclei. 相似文献
7.
The devil is in the detail: Nonadditive and context‐dependent plant population responses to increasing temperature and precipitation 下载免费PDF全文
Joachim P. Töpper Eric Meineri Siri L. Olsen Knut Rydgren Olav Skarpaas Vigdis Vandvik 《Global Change Biology》2018,24(10):4657-4666
In climate change ecology, simplistic research approaches may yield unrealistically simplistic answers to often more complicated problems. In particular, the complexity of vegetation responses to global climate change begs a better understanding of the impacts of concomitant changes in several climatic drivers, how these impacts vary across different climatic contexts, and of the demographic processes underlying population changes. Using a replicated, factorial, whole‐community transplant experiment, we investigated regional variation in demographic responses of plant populations to increased temperature and/or precipitation. Across four perennial forb species and 12 sites, we found strong responses to both temperature and precipitation change. Changes in population growth rates were mainly due to changes in survival and clonality. In three of the four study species, the combined increase in temperature and precipitation reflected nonadditive, antagonistic interactions of the single climatic changes for population growth rate and survival, while the interactions were additive and synergistic for clonality. This disparity affects the persistence of genotypes, but also suggests that the mechanisms behind the responses of the vital rates differ. In addition, survival effects varied systematically with climatic context, with wetter and warmer + wetter transplants showing less positive or more negative responses at warmer sites. The detailed demographic approach yields important mechanistic insights into how concomitant changes in temperature and precipitation affect plants, which makes our results generalizable beyond the four study species. Our comprehensive study design illustrates the power of replicated field experiments in disentangling the complex relationships and patterns that govern climate change impacts across real‐world species and landscapes. 相似文献
8.
Few studies have investigated the presence of shift work disorder (SWD) in the general community. We addressed many of the limitations in this literature and present new findings. SWD has been treated as an ‘all or none’ construct but we propose the need to consider the ‘severity’ of the disorder. Using random digit dialling, we randomly recruited 1163 participants. Participants completed an extensive battery of scales and questions concerning work, health and individual differences. Three questions based on the criteria from the International Classification for Sleep Disorders were used to categorise participants with SWD (n = 176). In addition, we asked participants whether SWD interfered with aspects of their life and high ratings were used to define severe shift work disorder (SSWD). The prevalence of SWD was 32.1% among night workers and 10.1% in day workers (p<.001). SSWD was present in 9.1% of night workers and 1.3% of day workers (p<.001). Adjusted logistic regression analyses found significant associations between SWD and night work (OR = 3.35, CI 2.19-5.12), weekly work hours (OR = 1.02, CI 1.00–1.04), short sleep (≤6 h; OR = 2.93, CI 1.94–4.41), languidity (OR = 1.11, CI 1.06–1.16) and resilience (OR = 0.56, CI 0.43–0.81). Night work, short sleep, languidity, and hypertension were significantly associated with SSWD. Overall, participants with SSWD slept 0.80 h less than other participants (p<.001). Night work, short sleep and languidity were associated with both SWD and SSWD. Day workers with SWD symptoms reported significantly shorter sleep duration, higher levels of languidity and worked longer working hours compared to day workers without SWD. 相似文献
9.
Algal Turbidity Hampers Ornament Perception,but Not Expression,in a Sex‐Role‐Reversed Pipefish 下载免费PDF全文
Josefin Sundin Gunilla Rosenqvist Siri Myhren Anders Berglund 《Ethology : formerly Zeitschrift fur Tierpsychologie》2016,122(3):215-225
Sexual ornaments are used both in intra‐ and intersexual contexts, and these signals have evolved to function in the particular habitat the animal is adapted to. Habitat characteristics may, however, change rapidly due to anthropogenic effects, sometimes at rates too fast for many organisms to adaptively respond. In aquatic ecosystems, eutrophication is currently changing chemical as well as visual properties of the environment. Algae blooms increase water turbidity, and the reduction of water transparency thus has the potential to alter visual ornaments and their perception. However, results are not congruent. Rather, algae turbidity may decrease, increase, or leave ornaments unaffected. The effect seems to depend on exposure time, condition, population and species. Here, we found that the perception of sexual signals, but not their expression, was hampered by turbidity in the sex‐role‐reversed pipefish Nerophis ophidion. In a laboratory experiment we found that female sexual ornaments (i.e., blue color markings and a skinfold) and fecundity was unaffected by turbidity. Male adaptive mate choice for larger females with large ornament was, however, hampered under turbid conditions, whereas in clear water males choose larger, more ornamented females. Thus, we show that water turbidity had no effect on signal expression but did hamper ornament perception and consequently randomized mate choice. 相似文献
10.