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11.
Magnusdottir A Stenmark P Flodin S Nyman T Kotenyova T Gräslund S Ogg D Nordlund P 《Proteins》2009,74(1):212-221
The PP2A serine/threonine phosphatase regulates a plethora of cellular processes. In the cell the predominant form of the enzyme is a heterotrimer, formed by a core dimer composed of a catalytic and a scaffolding subunit, which assemble together with one of a range of different regulatory B subunits. Here, we present the first structure of a free non-complexed B subunit, B56 gamma. Comparison with the recent structures of a heterotrimeric complex and the core dimer reveals several significant conformational changes in the interface region between the B56 gamma and the core dimer. These allow for an assembly scheme of the PP2A holoenzyme to be put forth where B56 gamma first complexes with the scaffolding subunit and subsequently binds to the catalytic subunit and this induces the formation of a binding site for the invariant C-terminus of the catalytic subunit that locks in the complex as a last step of assembly. 相似文献
12.
Differentiation-associated modulation of heparan sulfate structure and function in CaCo-2 colon carcinoma cells 总被引:3,自引:2,他引:1
Salmivirta M; Safaiyan F; Prydz K; Andresen MS; Aryan M; Kolset SO 《Glycobiology》1998,8(10):1029-1036
Heparan sulfate species expressed by different cell and tissue types differ
in their structural and functional properties. Limited information is
available on differences in regulation of heparan sulfate biosynthesis
within a single tissue or cell population under different conditions. We
have approached this question by studying the effect of cell
differentiation on the biosynthesis and function of heparan sulfate in
human colon carcinoma cells (CaCo-2). These cells undergo spontaneous
differentiation in culture when grown on semipermeable supports; the
differentiated cells show phenotypic similarity to small intestine
enterocytes. Metabolically labeled heparan sulfate was isolated from the
apical and basolateral media from cultures of differentiated and
undifferentiated cells. Compositional analysis of disaccharides, derived
from the contiguous N-sulfated regions of heparan sulfate, indicated a
greater proportion of 2-O- sulfated iduronic acid units and a smaller
amount of 6-O-sulfated glucosamine units in differentiated than in
undifferentiated cells. By contrast, the overall degree of sulfation, the
chain length and the size distribution of the N-acetylated regions were
similar regardless the differentiation status of the cells. The structural
changes were found to affect the binding of heparan sulfate to the long
isoform of platelet-derived growth factor A chain but not to fibroblast
growth factor 2. These findings show that heparan sulfate structures change
during cell differentiation and that heparan sulfate-growth factor
interactions may be affected by such changes.
相似文献
13.
Serglycin is the major proteoglycan in most hematopoietic cells, including
monocytes and macrophages. The monoblastic cell line U937-1 was used to
study the expression of serglycin during proliferation and differentiation.
In unstimulated proliferating U937-1 cells serglycin mRNA is
nonconstitutively expressed. The level of serglycin mRNA was found to
correlate with the synthesis of chondroitin sulfate proteoglycan (CSPG).
The U937-1 cells were induced to differentiate into different types of
macrophage-like cells by exposing the cells to PMA, RA, or VitD3. These
inducers of differentiation affected the expression of serglycin mRNA in
three different ways. The initial upregulation seen in the normally
proliferating cells was not observed in PMA treated cells. In contrast, RA
increased the initial upregulation, giving a reproducible six times
increase in serglycin mRNA level from 4 to 24 h of incubation, compared to
a four times increase in the control cells. VitD3 had no effect on the
expression of serglycin mRNA. The incorporation of (35S)sulfate into CSPG
decreased approximately 50% in all three differentiated cell types.
Further, the (35S)CSPGs expressed were of larger size in PMA treated cells
than controls, but smaller after RA treatment. This was due to the
expression of CSPGs, with CS-chains of 25 and 5 kDa in PMA and RA treated
cells, respectively, compared to 11 kDa in the controls. VitD3 had no
significant effect on the size of CSPG produced. PMA treated cells secreted
75% of the (35S)PGs expressed, but the major portion was retained in cells
treated with VitD3 or RA. The differences seen in serglycin mRNA levels,
the macromolecular properties of serglycin and in the PG secretion
patterns, suggest that serglycin may have different functions in different
types of macrophages.
相似文献
14.
Here we describe DNA sample preparation methods that allow the rapid, simultaneous generation of both single-stranded conformational polymorphism (SSCP) and heteroduplex DNA elements from a single sample in a single tube, which are suitable for direct injection into a capillary electrophoresis (CE) instrument with excellent sensitivity of genetic mutation detection. The p53 gene was used as a model DNA region for this study, which was performed on a high-throughput MegaBACE 96-capillary array electrophoresis instrument. We found that, contrary to the practice common in slab-gel SSCP analysis, denaturants such as formamide are incompatible with this novel technique because they result in homo- and heteroduplex peak broadening in CE (possibly as a result of incomplete dsDNA re-hybridization) that reduces the peak resolution and hence the sensitivity of mutation detection. We also have found that PCR buffers, which are typically used to suspend samples for slab-gel heteroduplex analysis (HA), but which are less suitable for CE because of the presence of extra salt that reduces the efficiency of electrokinetic injection, may be substituted with a 10 mM Tris-HCI buffer (pH 8.5). The use of this Tris-HCl buffer for sample preparation provides both a high sensitivity of mutation detection by tandem SSCP/HA and high efficiency ofelectrokinetic injection by CE. In a related study (published elsewhere), we have applied this optimized protocol to the screening of a set of 32 mutant DNA samples from p53 exons 7 and 8 and recorded 100% sensitivity of mutation detection for tandem CE-SSCP/HA, whereas each individual method yielded lower sensitivity on its own (93% for SSCP and 75% for HA). 相似文献
15.
Residuals for multinomial models 总被引:1,自引:0,他引:1
16.
SO Akarca-Dizakar H Aktuğ F Oltulu G Öktem A Yavaşoğlu E Açikgöz 《Biotechnic & histochemistry》2013,88(5):328-335
Diabetes mellitus (DM) affects many organs including kidney. Tyrosine kinase can cause hypoglycemia and sunitinib is an inhibitor of tyrosine kinase. We investigated the possible effects of sunitinib on the kidney of streptozotocin (STZ) induced type 1 diabetic mice. We used 28 CD 1 type male mice divided into four groups of seven. Type 1 diabetes was induced by injection of STZ. Group 1 was the untreated control. Group 2 comprised non-diabetic mice + sunitinib. Both groups 1 and 2 exhibited normal blood glucose levels. Group 3 comprised STZ treated diabetic mice + saline. Group 4 were diabetic mice + sunitinib treatment. Kidneys were removed after 8 weeks. The immunoreactivities of vimentin, E-cadherin and S100 were assessed. Immunostaining of vimentin, E-cadherin and S100 was located in both the glomeruli and tubules of the kidney. We found that the number of vimentin and E-cadherin positive glomeruli and tubules were increased after sunitinib treatment compared to saline treated diabetic mice. The number of vimentin labeled tubules was decreased in the sunitinib treated group compared to diabetic + saline groups. Differences in the number of S100 positive tubules and glomeruli between groups 3 and 4 were not statistically significant. The effect of sunitinib on experimental diabetic mice appears to be related to levels of vimentin, E-cadherin and S100 in the glomeruli and tubules of the kidney, and sunitinib may protect against renal damage from DM. 相似文献
17.
18.
Hyperinsulinemic, euglycemic clamp tests were performed on calves before and after clenbuterol treatment. Clenbuterol was given as 2 intramuscular injections with an interval of about 12 h. The dose used was 1 μg/kg b.w. The treatment resulted in increased plasma levels of insulin and glucose. The results of the clamp tests showed that clenbuterol induced a transient decrease in insulin sensitivity. Both insulin mediated glucose disposal (M), expressed as μmol/kg live b.w./min. and the M/I-index (M divided by the average insulin concentration at steady state) were significantly reduced after treatment. The effect of clenbuterol on carbohydrate metabolism seemed to be rather short-lived, since significant changes occurred only in animals treated 5-6 h prior to the test. According to the literature, the metabolic effects of clenbuterol have been studied only after the high doses used for growth promoting purposes. The results from the present study showed that similar changes occur also after doses at the therapeutic level. The hyperinsulinemic, euglycemic clamp test was considered to be a valuable tool for the study of insulin sensitivity in cattle. 相似文献
19.
Margret Bessadottir Mar Egilsson Eydis Einarsdottir Iris H. Magnusdottir Margret H. Ogmundsdottir Sesselja Omarsdottir Helga M. Ogmundsdottir 《PloS one》2012,7(12)
The lichen compound usnic acid (UA) is a lipophilic weak acid that acts as a proton shuttle and causes loss of mitochondrial inner membrane potential. In the current study we show that UA treatment induced the formation of autophagosomes in human cancer cells, but had minimal effects on normal human fibroblasts. However, autophagic flux was incomplete, degradation of autophagosomal content did not occur and acidification was defective. UA-treated cells showed reduced ATP levels and activation of AMP kinase as well as signs of cellular stress. UA is thus likely to trigger autophagosome formation both by energy depletion and stress conditions. Our findings indicate that the H+-shuttling effect of UA operates not only in mitochondria as previously shown, but also in lysosomes, and have implications for therapeutic manipulation of autophagy and pH-determined drug distribution. 相似文献
20.
Abstract. 1 The effect of larval rearing density on life-history parameters of Boettcherisca formosensis Kirner & Lopes (Sarcophagidae) was investigated. Increases in rearing density resulted in lowered larval survivorship, shortened larval development time and production of smaller, shorter-lived adults with reduced fecundity.
2. B. formosensis is larviparous. Average brood size was 17.5±1.0 (mean±M) larvae, which was much less than the average number of mature larvae inside gravid females. Females apparently produced a series of small broods, distributing their offspring over a number of carcasses.
3. Compared with the oviparous species Hemipyrellia ligurriens (Wiedemann) (Calliphoridae), B. formosensis adults were larger and longer-lived, with a longer larval development time but shorter larval feeding period. However, females had a shorter pre-reproductive period, were less fecund, and had a lower life time reproductive investment.
4. B. formosensis had lower relative performance (measured by the composite index of performance, r') than H. ligurriens over the larval rearing density range, and was more sensitive to increases in density. Although the r' values suggest that the sarcophagid may be a competitively inferior species, other features which are not included in the index (such as larvipary, short larval feeding period and spreading of offspring from a single brood among carcasses) may be of significant adaptive value to B. formosensis. 相似文献
2. B. formosensis is larviparous. Average brood size was 17.5±1.0 (mean±M) larvae, which was much less than the average number of mature larvae inside gravid females. Females apparently produced a series of small broods, distributing their offspring over a number of carcasses.
3. Compared with the oviparous species Hemipyrellia ligurriens (Wiedemann) (Calliphoridae), B. formosensis adults were larger and longer-lived, with a longer larval development time but shorter larval feeding period. However, females had a shorter pre-reproductive period, were less fecund, and had a lower life time reproductive investment.
4. B. formosensis had lower relative performance (measured by the composite index of performance, r') than H. ligurriens over the larval rearing density range, and was more sensitive to increases in density. Although the r' values suggest that the sarcophagid may be a competitively inferior species, other features which are not included in the index (such as larvipary, short larval feeding period and spreading of offspring from a single brood among carcasses) may be of significant adaptive value to B. formosensis. 相似文献