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Fiona Cuskin James E. Flint Tracey M. Gloster Carl Morland Arnaud Baslé Bernard Henrissat Pedro M. Coutinho Andrea Strazzulli Alexandra S. Solovyova Gideon J. Davies Harry J. Gilbert 《Proceedings of the National Academy of Sciences of the United States of America》2012,109(51):20889-20894
Noncatalytic carbohydrate binding modules (CBMs) are components of glycoside hydrolases that attack generally inaccessible substrates. CBMs mediate a two- to fivefold elevation in the activity of endo-acting enzymes, likely through increasing the concentration of the appended enzymes in the vicinity of the substrate. The function of CBMs appended to exo-acting glycoside hydrolases is unclear because their typical endo-binding mode would not fulfill a targeting role. Here we show that the Bacillus subtilis exo-acting β-fructosidase SacC, which specifically hydrolyses levan, contains the founding member of CBM family 66 (CBM66). The SacC-derived CBM66 (BsCBM66) targets the terminal fructosides of the major fructans found in nature. The crystal structure of BsCBM66 in complex with ligands reveals extensive interactions with the terminal fructose moiety (Fru-3) of levantriose but only limited hydrophobic contacts with Fru-2, explaining why the CBM displays broad specificity. Removal of BsCBM66 from SacC results in a ∼100-fold reduction in activity against levan. The truncated enzyme functions as a nonspecific β-fructosidase displaying similar activity against β-2,1– and β-2,6–linked fructans and their respective fructooligosaccharides. Conversely, appending BsCBM66 to BT3082, a nonspecific β-fructosidase from Bacteroides thetaiotaomicron, confers exolevanase activity on the enzyme. We propose that BsCBM66 confers specificity for levan, a branched fructan, through an “avidity” mechanism in which the CBM and the catalytic module target the termini of different branches of the same polysaccharide molecule. This report identifies a unique mechanism by which CBMs modulate enzyme function, and shows how specificity can be tailored by integrating nonspecific catalytic and binding modules into a single enzyme. 相似文献
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Heterogeneity of the response of chronic lymphocytic leukemia cells to phorbol ester 总被引:3,自引:0,他引:3
The ability of the tumor promotor 12-0-tetradecanoylphorbol 13-acetate (TPA) to induce differentiation of leukemic cells was studied in 10 cases of chronic lymphocytic leukemia (CLL). An increase in modal volume and an enhancement of the capacity of te leukemic cells to stimulate in mixed lymphocyte reaction (MLR) was seen in the majority of cases. A significant increase in Ia expression was observed upon culture of leukemic cells with TPA in 6 of the 10 cases; 5 of these cases also showed an induction of cytoplasmic IgM production. Correlations between the phenotypic markers of the leukemic cells and their ability to respond to TPA were evaluated. CLL cells with low amounts to surface Ig. a volume less than or equal to 165 fl. and relatively low la expression responded well to TPA. Cells with bright surface Ig. a volume greater than or equal to 178 fl. and elevated amounts of Ia responded poorly to TPA. These results suggest that differences in the response of B leukemic cells to TPA reflect the underlying heterogeneity of the leukemic cells and might be correlated with their stage of maturation. 相似文献
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A quality assurance programme was used to evaluate community and primary care based preschool surveillance using the National Child Health Computer System in 40 examination centres. Quarterly reports were generated from returns from clinical medical officers and general practitioners to list non-attenders, uptake, and timeliness for the four preschool checks. These provided rapid and comparative feedback on personal performance for participating health professionals and led to marked rises in recorded timeliness and uptake against preset targets. Pre-existing uptake was highest at the 6 week check with least overall improvement. Greatest improvements occurred at the 18 month health visitor check but, in general, results plateaued when the programme had been in use for 12 to 18 months. Particular problems such as data legibility and mobile populations were identified and solutions formulated. It is postulated that improvements in performance were due to enhanced professional motivation as no other factors changed. This system provides a valuable contribution in the light of changing patterns of service provision. 相似文献
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No Effect of Acute Ethanol Administration on Hepatic Protein Synthesis and Export in the Rat in Vivo
Bengt Wallin MSc Atle Bessesen Anne-Marie Fikke BSc Jarle Aarbakke MD Jorg Morland MD 《Alcoholism, clinical and experimental research》1984,8(2):191-195
Ethanol was administered as a single p.o. dose (2.88 g X kg-1) to male rats (220-265 g body weight) to give blood alcohol concentrations of 40-50 mM for the following 3 hr. Controls were given isoenergetic amounts of either sucrose or lipid. Liver protein synthetic rates were measured during a 20 min interval at the end of the 3 hr period following the administration of diets. Although ethanol caused a 32% reduction of the incorporation of labelled valine into liver protein compared to the sucrose group during the 20 min interval, no such reduction was found when the synthetic rate of stationary liver protein was calculated (182 vs. 214 (not significant) pmol X mg protein-1 X min-1) for same interval. There was no difference between the ethanol and lipid group with regard to either incorporation or synthetic rates. Incorporation of valine into plasma proteins was reduced in the ethanol group compared to the sucrose group, but not compared to the lipid control group, again demonstrating no ethanol-specific effect. When the incorporation into plasma proteins was divided by the specific radioactivity of valyl-tRNA at 20 min, the difference between the ethanol and the sucrose group disappeared. The fraction of newly synthesized proteins exported to the plasma measured 40 min after the injection of labeled valine, was equal in all three treatment groups. It was concluded that acute administration of ethanol has no consistent effect on liver protein synthesis and secretion in vivo. 相似文献