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Proteoglycans were separated by high-performance liquid chromatography (HPLC), using two coupled Aquapore columns containing glycerylpropylsilane groups covalently linked to large-pore (50–100 nm) silica spheres. This two-column HPLC system was effective in separating cartilage proteoglycan aggregates and monomers, without altering their biochemical integrity. This system was also effective in resolving small amounts of isotopically labeled proteoglycans synthesized by cultured mammalian cells. The small sample size, short analysis time, and high reproducibility represent improvements in the study of proteoglycans over conventional soft-gel chromatography. 相似文献
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Sterol Composition and Accumulation in Glycine max (L.) Merr Leaves under Different Filtered Sunlight Conditions
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Izzo R 《Plant physiology》1981,67(6):1073-1077
Soybean plants (Merr) were grown in the field in three plots. Sixteen days after sowing, two plots were covered with blue and red polyvinylchloride filters (0.45 millimeter thick) and one remained uncovered as control. Leaves of all plots were analyzed for total, free, esterified, and glycosidic sterols at two successive stages of plant growth (flowering and podripening). 相似文献
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Drabløs F Feyzi E Aas PA Vaagbø CB Kavli B Bratlie MS Peña-Diaz J Otterlei M Slupphaug G Krokan HE 《DNA Repair》2004,3(11):1389-1407
Alkylation lesions in DNA and RNA result from endogenous compounds, environmental agents and alkylating drugs. Simple methylating agents, e.g. methylnitrosourea, tobacco-specific nitrosamines and drugs like temozolomide or streptozotocin, form adducts at N- and O-atoms in DNA bases. These lesions are mainly repaired by direct base repair, base excision repair, and to some extent by nucleotide excision repair (NER). The identified carcinogenicity of O(6)-methylguanine (O(6)-meG) is largely caused by its miscoding properties. Mutations from this lesion are prevented by O(6)-alkylG-DNA alkyltransferase (MGMT or AGT) that repairs the base in one step. However, the genotoxicity and cytotoxicity of O(6)-meG is mainly due to recognition of O(6)-meG/T (or C) mispairs by the mismatch repair system (MMR) and induction of futile repair cycles, eventually resulting in cytotoxic double-strand breaks. Therefore, inactivation of the MMR system in an AGT-defective background causes resistance to the killing effects of O(6)-alkylating agents, but not to the mutagenic effect. Bifunctional alkylating agents, such as chlorambucil or carmustine (BCNU), are commonly used anti-cancer drugs. DNA lesions caused by these agents are complex and require complex repair mechanisms. Thus, primary chloroethyl adducts at O(6)-G are repaired by AGT, while the secondary highly cytotoxic interstrand cross-links (ICLs) require nucleotide excision repair factors (e.g. XPF-ERCC1) for incision and homologous recombination to complete repair. Recently, Escherichia coli protein AlkB and human homologues were shown to be oxidative demethylases that repair cytotoxic 1-methyladenine (1-meA) and 3-methylcytosine (3-meC) residues. Numerous AlkB homologues are found in viruses, bacteria and eukaryotes, including eight human homologues (hABH1-8). These have distinct locations in subcellular compartments and their functions are only starting to become understood. Surprisingly, AlkB and hABH3 also repair RNA. An evaluation of the biological effects of environmental mutagens, as well as understanding the mechanism of action and resistance to alkylating drugs require a detailed understanding of DNA repair processes. 相似文献
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The peregrine falcon (Falco peregrinus) population in southern Scandinavia was almost extinct in the 1970’s. A successful reintroduction project was launched in
1974, using captive breeding birds of northern and southern Scandinavian, Finnish and Scottish origin. We examined the genetic
structure in the pre-bottleneck population using eleven microsatellite markers and compared the data with the previously genotyped
captive breeding population and contemporary wild population. Museum specimens between 53 and 130 years old were analyzed.
Despite an apparent loss of historical genetic diversity, the contemporary population shows a relatively high level of genetic
variation. Considerable gene introgression from captive breeding stock used to repopulate the former range of southern Scandinavian
peregrines may have altered the genetic composition of this population. Both the historical and contemporary northern and
southern Scandinavian populations are genetically differentiated. The reintroduction project implemented in the region and
the use of non-native genetic stock likely prevented the southern Scandinavian population from extinction and thus helped
maintain the level of genetic diversity and prevent inbreeding depression. The population is rapidly increasing in numbers
and range and shows no indication of reduced fitness or adaptive capabilities in the wake of the severe bottleneck and the
reintroduction. 相似文献
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MH Daleke AD van der Woude AH Parret R Ummels AM de Groot D Watson SR Piersma CR Jiménez J Luirink W Bitter EN Houben 《The Journal of biological chemistry》2012,287(38):31939-31947
Mycobacteria use the dedicated type VII protein secretion systems ESX-1 and ESX-5 to secrete virulence factors across their highly hydrophobic cell envelope. The substrates of these systems include the large mycobacterial PE and PPE protein families, which are named after their characteristic Pro-Glu and Pro-Pro-Glu motifs. Pathogenic mycobacteria secrete large numbers of PE/PPE proteins via the major export pathway, ESX-5. In addition, a few PE/PPE proteins have been shown to be exported by ESX-1. It is not known how ESX-1 and ESX-5 recognize their cognate PE/PPE substrates. In this work, we investigated the function of the cytosolic protein EspG(5), which is essential for ESX-5-mediated secretion in Mycobacterium marinum, but for which the role in secretion is not known. By performing protein co-purifications, we show that EspG(5) interacts with several PPE proteins and a PE/PPE complex that is secreted by ESX-5, but not with the unrelated ESX-5 substrate EsxN or with PE/PPE proteins secreted by ESX-1. Conversely, the ESX-1 paralogue EspG(1) interacted with a PE/PPE couple secreted by ESX-1, but not with PE/PPE substrates of ESX-5. Furthermore, structural analysis of the complex formed by EspG(5) and PE/PPE indicates that these proteins interact in a 1:1:1 ratio. In conclusion, our study shows that EspG(5) and EspG(1) interact specifically with PE/PPE proteins that are secreted via their own ESX systems and suggests that EspG proteins are specific chaperones for the type VII pathway. 相似文献
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