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51.
The structure of human collagen type IX and its organization in fetal and infant cartilage fibrils 总被引:1,自引:0,他引:1
P Bruckner M Mendler B Steinmann S Huber K H Winterhalter 《The Journal of biological chemistry》1988,263(32):16911-16917
Human collagen type IX was isolated from the media of organ cultures of fetal or infant hyaline cartilage. It consisted of three distinct, disulfide-bonded polypeptides of 115, 84, and 72 kDa, respectively. Digestion with chondroitinase ABC reduced the apparent molecular mass of the 115-kDa chain to about 65 kDa demonstrating that also human collagen type IX is a proteoglycan. In the electron microscope, the molecule had a rigid rod-like structure with characteristic kinks and with a globular domain at one end. Digestion of human collagen type IX with pepsin leads to somewhat heterogeneous fragments. Affinity-purified antibodies to the mixture of fragments specifically reacted with the fragment HMW without cross-reaction with chicken HMW. LMW of both species were recognized to the same low extent. Mechanically generated fibril fragments from human fetal cartilage were heterogeneous in diameter. Significantly, they could be immunostained for collagen type IX in a D-periodic pattern and regardless of the fibril diameter. Some fibrils were poorly labeled, again independently of the diameter. Therefore, the role of collagen type IX in cartilage probably is not to control directly the lateral growth during fibrillogenesis but rather to stabilize the fibril network. 相似文献
52.
Upstream G gamma-globin and downstream beta-globin sequences required for stage-specific expression in transgenic mice. 总被引:16,自引:6,他引:16
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The human G gamma-globin and beta-globin genes are expressed in erythroid cells at different stages of human development, and previous studies have shown that the two cloned genes are also expressed in a differential stage-specific manner in transgenic mice. The G gamma-globin gene is expressed only in murine embryonic erythroid cells, while the beta-globin gene is active only at the fetal and adult stages. In this study, we analyzed transgenic mice carrying a series of hybrid genes in which different upstream, intragenic, or downstream sequences were contributed by the beta-globin or G gamma-globin gene. We found that hybrid 5'G gamma/3'beta globin genes containing G gamma-globin sequences upstream from the initiation codon were expressed in embryonic erythroid cells at levels similar to those of an intact G gamma-globin transgene. In contrast, beta-globin upstream sequences were insufficient for expression of 5'beta/3'G gamma hybrid globin genes or a beta-globin-metallothionein fusion gene in adult erythroid cells. However, beta-globin downstream sequences, including 212 base pairs of exon III and 1,900 base pairs of 3'-flanking DNA, were able to activate a 5'G gamma/3'beta hybrid globin gene in fetal and adult erythroid cells. These experiments suggest that positive regulatory elements upstream from the G gamma-globin and downstream from the beta-globin gene are involved in the differential expression of the two genes during development. 相似文献
53.
54.
Lipid raft targeting of hematopoietic protein tyrosine phosphatase by protein kinase C theta-mediated phosphorylation
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Nika K Charvet C Williams S Tautz L Bruckner S Rahmouni S Bottini N Schoenberger SP Baier G Altman A Mustelin T 《Molecular and cellular biology》2006,26(5):1806-1816
Protein kinase C theta (PKC theta) is unique among PKC isozymes in its translocation to the center of the immune synapse in T cells and its unique downstream signaling. Here we show that the hematopoietic protein tyrosine phosphatase (HePTP) also accumulates in the immune synapse in a PKC theta-dependent manner upon antigen recognition by T cells and is phosphorylated by PKC theta at Ser-225, which is required for lipid raft translocation. Immune synapse translocation was completely absent in antigen-specific T cells from PKC theta-/- mice. In intact T cells, HePTP-S225A enhanced T-cell receptor (TCR)-induced NFAT/AP-1 transactivation, while the acidic substitution mutant was as efficient as wild-type HePTP. We conclude that HePTP is phosphorylated in the immune synapse by PKC theta and thereby targeted to lipid rafts to temper TCR signaling. This represents a novel mechanism for the active immune synapse recruitment and activation of a phosphatase in TCR signaling. 相似文献
55.
Christian Kampichler and Alexander Bruckner 《Biological reviews of the Cambridge Philosophical Society》2009,84(3):375-389
Litterbags have been utilized in soil ecology for about 50 years. They are useful because they confine organic material and thus enable the study of decomposition dynamics (mass loss and/or nutrient loss through time, colonization by soil biota) in situ, i.e. under field conditions. Researchers can easily restrict or permit access to certain size classes of soil fauna to determine their contribution to litter mass loss by choosing adequate mesh size or applying specific biocides. In particular, the mesofauna has received much attention since it comprises two very abundant and diverse microarthropod groups, the Collembola (springtails) and Acari (mites). We comprehensively searched the literature from the mid‐1960s to the end of 2005 for reports on litterbag experiments investigating the role of microarthropods in terrestrial decomposition. Thirty papers reporting 101 experiments satisfied our selection criteria and were included in the database. Our meta‐analysis revealed that microarthropods have a moderate but significant effect on mass loss. We discuss in detail the interactions of the microarthropod effect with study characteristics such as experimental design (e.g. number of bags, duration of experiment), type of exposed organic matter, climatic zone and land use of the study site. No publication bias was detected; however, we noticed a significant decrease in the microarthropod effect with publication year, indicating that, in the first decades of litterbag use, soil zoologists may have studied “promising” sites with a higher a priori probability of positive microarthropod effects on litter mass loss. A general weakness is that the treatments differ not only with respect to the presence or absence of microarthropods, but also with regard to mesh size (small to exclude microarthropods, wide to permit their access) or presence (to exclude microarthropods) and absence (to permit their access) of an insecticide. Consequently, the difference between the decomposition rates in the treatments is not a pure microarthropod effect but will be influenced by the additive effects of mesh size and insecticide. The relative contribution of the “true” microarthropod effect remains unknown without additional treatments controlling for the differential mesh size/insecticide effect. A meta‐analysis including only those studies using different mesh size and for which the data were corrected by subtracting an estimated mesh size effect based on data from the literature yielded a significantly negative microarthropod effect on litter decomposition. These results cast doubt on the widely accepted hypothesis that microarthropods generally exert a positive effect on litter mass loss. We conclude that after 40 years of litterbag studies our knowledge on the role of microarthropods in litter mass loss remains limited and that the inclusion of a third treatment in future studies is a promising way to retain litterbags as a meaningful tool of soil biological studies. 相似文献
56.
Zilun Hu Xiangjun Jiang Charles F. Albright Nilsa Graciani Eddy Yue Mingzhu Zhang Shu-Yun Zhang Robert Bruckner Melody Diamond Randine Dowling Maria Rafalski Swamy Yeleswaram George L. Trainor Steven P. Seitz Wei Han 《Bioorganic & medicinal chemistry letters》2010,20(3):853-856
To selectively target doxorubicin (Dox) to tumor tissue and thereby improve the therapeutic index and/or efficacy of Dox, matrix metalloproteinases (MMP) activated peptide–Dox prodrugs were designed and synthesized by coupling MMP-cleavable peptides to Dox. Preferred conjugates were good substrates for MMPs, poor substrates for neprilysin, an off-target proteinase, and stable in blood ex vivo. When administered to mice with HT1080 xenografts, conjugates, such as 19, preferentially released Dox in tumor relative to heart tissue and prevented tumor growth with less marrow toxicity than Dox. 相似文献
57.
Suassuna Tde M Bruckner H de Carvalho R Borém A 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2003,106(2):298-302
Self-incompatibility in passionfruit was studied in families originated from crosses among plants that presented differences in reciprocal crosses. The three families, obtained by crossing S(3) plants, exhibited one incompatible group; no reciprocal differences were observed. The phenotype of the families was the same as the parent plants, S(3). These results suggest the presence of a gene ( G), gametophytic in its action, associated to the sporophytic gene S, modifying the incompatibility reaction in passionfruit. The reciprocal difference exhibited in the crosses among the parents could be explained as a matching between plants homozygous for S, but homozygous and heterozygous for G. Actually this would be a partially compatible cross, not detectable when the evaluation is done based on fruit set data. As the family originated from this kind of cross is homozygous for S and heterozygous for G, no reciprocal differences are expected, and the phenotype should be the same as the parental plants, as observed in the present work. 相似文献
58.
Hambrock HO Nitsche DP Hansen U Bruckner P Paulsson M Maurer P Hartmann U 《The Journal of biological chemistry》2003,278(13):11351-11358
SC1, a member of the BM-40 family of extracellular matrix proteins, was recombinantly expressed in a eukaryotic expression system. The full-length protein as well as truncated versions were purified to homogeneity under non-denaturing conditions. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry of full-length SC1 revealed a mass of 87.8 kDa of which 16.8 kDa is contributed by posttranslational modifications. In electron microscopy, after negative staining, SC1 was revealed as a globule attached to a thread-like structure. A calcium dependence of the SC1 conformation could be demonstrated by fluorescence spectroscopy. In the extracellular matrix of cultured osteosarcoma cells SC1 was found associated with collagen I-containing fibrils, and binding of SC1 to reconstituted collagen I fibrils could be demonstrated by immunogold labeling and electron microscopy. SC1 showed a broad expression in a variety of tissues. 相似文献
59.
M. D. Gordon R. J. Ezzell N. I. Bruckner J. M. Ascenzi 《Journal of industrial microbiology & biotechnology》1994,13(2):77-82
Summary Several ,-unsaturated and aromatic aldehydes were evaluated for antimicrobial activity usingMycobacterium bovis as the test strain. Activity of most of the compounds was determined in the presence and absence of 2% glutaraldehyde. Several compounds highly active against this organism, e.g. 2-pentenal, benzaldehyde, ando-phthalaldehyde showed rapid kill of >105 CFU ml–1 in 5 min. Activity of ,-unsaturated compounds substituted in the 1 position showed increasing activity with increasing chain length. Of the aromatic aldehydes tested, benzaldehyde andp-dimethylamino benzaldehyde showed little activity alone, but when combined with 2% glutaraldehyde showed increased activity. Substituents added to the benzaldehyde ring (nitro, chloro, methyl, and methoxy) all detracted from the synergism, but still showed increased activity over the activity of 2% glutaraldehyde. The same affect was noted with disubstituted benzaldehyde compounds but not with substitutedo-phthaladehyde (2-formylformaldehyde). 相似文献
60.
The precursor-specific aminopropeptide of bovine type III procollagen is a strong immunogen in rabbits, guinea pigs and mice and induces antibodies which do not cross-react with type I procollagen. The antibody response is regulated by immune response genes associated with the major histocompatibility complex. Major antigenic determinants were found in the compact, non-collagenous domain (fragment Col 1) located at the N terminus of the aminopropeptide and were destroyed by reduction of disulfide bonds. Minor antigenic determinants independent of disulfide bonds also exist in fragment Col 1 and could be localized on a distinct tryptic peptide. Fragment Col 1 showed a lower affinity for antibody when compared with the intact aminopropeptide which causes a non-parallel shift in radioimmuno-inhibition profiles. Monovalent antibody fragments showed an average tenfold reduction in affinity constant and failed to distinguish between aminopropeptide and fragment Col 1. This indicates that the stronger binding of bivalent antibody by the triple-stranded aminopropeptide is due to multiple interactions with both antibody binding sites which are lost for a single-stranded antigen (Col 1) or with monovalent antibody fragments. 相似文献