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“The flaviviral nonstructural 3 protease (NS3pro) is essential for virus replication and is therefore a pharmaceutically relevant target to fight Dengue and West Nile virus (WNV). NS3pro is a chymotrypsin- like serine protease which requires a polypeptide cofactor (NS2B) for activation. Recent X-ray crystallography studies have led to the suggestion that the substrate binds to the two-component SHP099 mw NS2B-NS3pro enzyme by an induced-fit mechanism. Here, multiple explicit water molecular dynamics simulations of the WNV NS2B-NS3pro enzyme show that the active conformation of the NS2B cofactor ( in which its beta-loop is part of the substrate binding site) is stable
over a 50-ns time scale even in the absence of the inhibitor. The partial and reversible opening of the NSB2 beta-loop and its correlated motion with an adjacent NS3pro loop, both observed in the simulations started from the active conformation, are likely to facilitate substrate binding and product release. Moreover, in five CX-6258 purchase of eight simulations without inhibitor ( started from two X-ray structures both with improperly formed oxyanion hole) the Thr132-Gly133 peptide bond flips spontaneously thereby promoting the formation of the catalytically competent oxyanion hole, which then stays stable until the end of the runs. The
simulation results provide evidence at atomic level of detail that the substrate binds to the NS2B-NS3pro enzyme by conformational selection, rather than induced-fit mechanism.”
“Background The interest in neonatal screening for lysosomal storage disorders has increased substantially because of newly developed enzyme replacement Selleckchem NU7026 therapies, the need for early diagnosis, and technical advances. We tested for Gaucher’s disease, Pompe’s disease, Fabry’s disease, and Niemann-Pick disease types A and B in an anonymous prospective nationwide screening study that included genetic
mutation analysis to assess the practicality and appropriateness of including these disorders in neonatal screening panels.
Methods Specimens from dried blood spots of 34 736 newborn babies were collected consecutively from January, 2010 to July, 2010, as part of the national routine Austrian newborn screening programme. Anonymised samples were analysed for enzyme activities of acid beta-glucocerebrosidase, alpha-galactosidase, alpha-glucosidase, and acid sphingomyelinase by electrospray ionisation tandem mass spectrometry. Genetic mutation analyses were done in samples with suspected enzyme deficiency.
Findings All 34 736 samples were analysed successfully by the multiplex screening assay. Low enzyme activities were detected in 38 babies. Mutation analysis confirmed lysosomal storage disorders in 15 of them. The most frequent mutations were found for Fabry’s disease (1 per 3859 births), followed by Pompe’s disease (1 per 8684), and Gaucher’s disease (1 per 17 368).