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81.
Expression of SPARC during development of the chicken chorioallantoic membrane: evidence for regulated proteolysis in vivo. 总被引:5,自引:0,他引:5 下载免费PDF全文
M L Iruela-Arispe T F Lane D Redmond M Reilly R P Bolender T J Kavanagh E H Sage 《Molecular biology of the cell》1995,6(3):327-343
SPARC is a secreted glycoprotein that has been shown to disrupt focal adhesions and to regulate the proliferation of endothelial cells in vitro. Moreover, peptides resulting from the proteolysis of SPARC exhibit angiogenic activity. Here we describe the temporal synthesis, turnover, and angiogenic potential of SPARC in the chicken chorioallantoic membrane. Confocal immunofluorescence microscopy revealed specific expression of SPARC protein in endothelial cells, and significantly higher levels of SPARC were observed in smaller newly formed blood vessels in comparison to larger, developmentally older vessels. SPARC mRNA was detected at the earliest stages of chorioallantoic membrane morphogenesis and reached maximal levels at day 13 of embryonic development. Interestingly, steady-state levels of SPARC mRNA did not correlate directly with protein accumulation; moreover, the protein appeared to undergo limited degradation during days 10-15. Incubation of [125I]-SPARC with chorioallantoic membranes of different developmental ages confirmed that extracellular proteolysis occurred during days 9-15, but not at later stages (e.g., days 17-21). Comparison of peptides produced by incubation with chorioallantoic membranes with those generated by plasmin showed an identical pattern of proteolysis. Plasmin activity was present throughout development, and in situ zymography identified sites of plasminogen activator activity that corresponded to areas exhibiting high levels of SPARC expression. Synthetic peptides from a plasmin-sensitive region of SPARC, between amino acids 113-130, stimulated angiogenesis in the chorioallantoic membrane in a dose-dependent manner; in contrast, intact SPARC was inactive in similar assays. We have shown that SPARC is expressed in endothelial cells of newly formed blood vessels in a manner that is both temporally and spatially restricted. Between days 9 and 15 of chorioallantoic membrane development, the protein undergoes proteolytic cleavage that is mediated, in part, by plasmin. SPARC peptides released specifically by plasmin induce angiogenesis in vivo. We therefore propose that SPARC acts as an intrinsic regulator of angiogenesis in vivo. 相似文献
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Anne Imberty Florence Casset Colin V. Gegg Marilynn E. Etzler Serge Pérez 《Glycoconjugate journal》1994,11(5):400-413
The three-dimensional structure ofDolichos biflorus seed lectin has been constructed using five legume lectins for which high resolution crystal structures were available. The validity of the resulting model has been thoroughly investigated. Final structure optimization was conducted for the lectin complexed with GalNAc, providing thereby the first three-dimensional structure of lectin/GalNAc complex. The role of theN-acetyl group was clearly evidenced by the occurrence of a strong hydrogen bond between the protein and the carbonyl oxygen of the carbohydrate and by hydrophobic interaction between the methyl group and aromatic amino acids. Since the lectin specificity is maximum for the Forssman disaccharide GalNAc(1–3)GalNAc-O-Me and the blood group A trisaccharide GalNAc(1–3)[Fuc(1–2)]Gal-O-Me, the complexes with these oligosaccharides have been also modelled. 相似文献
84.
A lectin preparation obtained fromTetracarpidium conophorum (Nigerian walnut) by affinity chromatography of seed extracts on lactose-agarose has been shown to contain two components by gel filtration on Sephadex G150. The larger componentTetracarpidium conophorum agglutinin I (TCAI) is a disulphide-bonded 70 kDa homodimer whereas the second component TCAII is a 34 kDa monomeric protein. Amino terminal aminoacid sequencing shows identity in TCAI and TCAII for the first fifteen residues after which the sequences diverge. TheN-terminal sequences of TCAI and TCAII show identity with sequences in the B-chains of ricin andRicinus communis agglutinin I (RCAI) in eleven of the initial fifteen residues. Thereafter TCAI appears to be homologous to the ricin B chain whereas TCAII is more homologous with the B chain of RCAI. A limited screening of the carbohydrate-binding specificity of TCAII by affinity chromatography of defined oligosaccharides on TCAII Sepharose columns shows that the binding specificity reported earlier for affinity purifiedTetracarpidium conophorum isolectins (Sato S, Animashaun T, Hughes RC (1991)J Biol Chem
266:11485–94) reflects the binding properties of TCAII which is the major isolectin in unfractionated lectin preparations. 相似文献
85.
Guido Barbujani Andrea Pilastro Silvia De Domenico Colin Renfrew 《American journal of physical anthropology》1994,95(2):137-154
The hypothesis that both genetic and linguistic similarities among Eurasian and North African populations are due to demic diffusion of neolithic farmers is tested against a wide database of allele frequencies. Demic diffusion of farming and languages from the Near East should have determined clines in areas defined by linguistic criteria; the alternative hypothesis of cultural transmission does not predict clines. Spatial autocorrelation analysis shows significant gradients in three of the four linguistic families supposedly affected by neolithic demic diffusion; the Afroasiatic family is the exception. Many such gradients are not observed when populations are jointly analyzed, regardless of linguistic classification. This is incompatible with the hypothesis that major cultural transformations in Eurasia (diffusion of related languages and spread of agriculture) took place without major demographic changes. The model of demic diffusion seems therefore to provide a mechanism explaining coevolution of linguistic and biological traits in much of the Old World. Archaeological, linguistic, and genetic evidence agree in suggesting a multidirectional process of gene flow from the Near East in the neolithic. However, the possibility should be envisaged that some allele frequency patterns can predate the neolithic and depend on the initial spread of Homo sapiens sapiens from Africa into Eurasia. © 1994 Wiley-Liss, Inc. 相似文献
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May J. Reed Robert B. Vernon Itamar B. Abrass E. Helene Sage 《Journal of cellular physiology》1994,158(1):169-179
Fibroblasts have a major role in the synthesis and reorganization of extracellular matrix that occur during wound repair. An impaired biosynthetic or functional response of these cells to stimulation by growth factors might contribute to the delayed wound healing noted in aging. We, therefore, compared the responses of dermal fibroblasts from young and elderly individuals (26, 29, 65, 89, 90, and 92 years of age) to transforming growth factor-β1 (TGF-β1) with respect to: (1) the synthesis of type I collagen and SPARC (two extracellular matrix proteins that are highly expressed by dermal fibroblasts during the remodeling phase of wound repair) and (2) the contraction of collagen gels, an in vitro assay of wound contraction. With the exception of one young donor, all cultures exposed for 44 hours to 10 ng/ml TGF-β1 exhibited a 1.6- to 5.5-fold increase in the levels of secreted type 1 collagen and SPARC, relative to untreated cultures, and exhibited a 2.0- to 6.2-fold increase in the amounts of the corresponding mRNAs. Moreover, the dose-response to TGF-β1 (0.1–10 ng/ml), as determined by synthesis of type I collagen and SPARC mRNA, was as vigorous in cells from aged donors as in cells from a young donor. In assays of collagen gel contraction, fibroblasts from all donors were stimulated to a similar degree by 10 ng/ml TGF-β1. In conclusion, cells from both young and aged donors exhibited similar biosynthetic and contractile properties with exposure to TGF-β1. It therefore appears that the impaired wound healing noted in the aged does not result from a failure of their dermal fibroblasts to respond to this cytokine. © 1994 Wiley-Liss, Inc. 相似文献
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Acclimation of photosynthesis to increasing atmospheric CO2: The gas exchange perspective 总被引:9,自引:0,他引:9
Rowan F. Sage 《Photosynthesis research》1994,39(3):351-368
The nature of photosynthetic acclimation to elevated CO2 is evaluated from the results of over 40 studies focusing on the effect of long-term CO2 enrichment on the short-term response of photosynthesis to intercellular CO2 (the A/Ci response). The effect of CO2 enrichment on the A/Ci response was dependent on growth conditions, with plants grown in small pots (< 5 L) or low nutrients usually exhibiting a reduction of A at a given Ci, while plants grown without nutrient deficiency in large pots or in the field tended to exhibit either little reduction or an enhancement of A at a given Ci following a doubling or tripling of atmospheric CO2 during growth. Using theoretical interpretations of A/Ci curves to assess acclimation, it was found that when pot size or nutrient deficiency was not a factor, changes in the shape of A/Ci curves which are indicative of a reallocation of resources within the photosynthetic apparatus typically were not observed. Long-term CO2 enrichment usually had little effect or increased the value of A at all Ci. However, a minority of species grown at elevated CO2 exhibited gas exchange responses indicative of a reduced amount of Rubisco and an enhanced capacity to metabolize photosynthetic products. This type of response was considered beneficial because it enhanced both photosynthetic capacity at high CO2 and reduced resource investment in excessive Rubisco capacity. The ratio of intercellular to ambient CO2 (the Ci/Ca ratio) was used to evaluate stomatal acclimation. Except under water and humidity stress, Ci/Ca exhibited no consistent change in a variety of C3 species, indicating no stomatal acclimation. Under drought or humidity stress, Ci/Ca declined in high-CO2 grown plants, indicating stomata will become more conservative during stress episodes in future high CO2 environments.Abbreviations A
net CO2 assimilation rate
- Ci (Ca)
intercellular (ambient) partial pressure of CO2
- operational Ci
intercellular partial pressure of CO2 at a given ambient partial pressure of CO2
- gs
stomatal conductance
- normal CO2
current atmospheric mole fraction of CO2 (330 to 355 mol mol–1)
- Rubisco
ribulose-1,5-bisphosphate carboxylase/oxygenase 相似文献