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Dr. Vlastimil Jirasko Dr. Nils-Alexander Lakomek Dr. Susanne Penzel Dr. Marie-Laure Fogeron Prof. Dr. Ralf Bartenschlager Prof. Dr. Beat H. Meier Dr. Anja Böckmann 《Chembiochem : a European journal of chemical biology》2020,21(10):1453-1460
Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D 1H-detected 1H,15N and 3D 1H,13C,15N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies. 相似文献
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Document caching and connection caching are extensively studied problems. In document caching, one has to maintain caches
containing documents accessible in a network. In connection caching, one has to maintain a set of open network connections
that handle data transfer. Previous work investigated these two problems separately while in practice the problems occur together:
In order to load a document, one has to establish a connection between network nodes if the required connection is not already
open. In this paper we present the first study that integrates document and connection caching. We first consider a very basic
model in which all documents have the same size and the cost of loading a document or establishing a connection is equal to
1. We present deterministic and randomized online algorithms that achieve nearly optimal competitive ratios unless the size
of the connection cache is extremely small. We then consider general settings where documents have varying sizes. We investigate
a FAULT model in which the loading cost of a document is 1 as well as a BIT model in which the loading cost is equal to the
size of the document. 相似文献
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The effects of oxidation/reduction regeneration treatments, with and without 1,2-dichloropropane present as a chlorinating agent, on the structure of Pt(3%)–Sn(4.5%)/Al2O3 catalysts have been correlated with selectivities for butane/H2 reactions. Particles of Pt0 fin Cl-free catalysts were partly covered by Sn0, but retained exposed ensembles of Pt atoms which were active for isomerisation, hydrogenolysis and dehydrogenation reactions, the latter becoming dominant at high reaction temperatures. Coking reduced Pt ensemble size and, hence, also favoured high selectivities for dehydrogenation as hydrogenolysis and isomerisation sites became poisoned. In contrast, the addition of 1,2-dichloropropane in an oxychlorination step before reduction promoted 1:1 Pt0–Sn0 alloy formation after reduction, the proportion of the total Pt in alloy being enhanced by increasing 1,2-dichloropropane concentration and oxychlorination temperature. The alloy surfaces were inactive for isomerisation and hydrogenolysis reactions, giving dehydrogenation as the sole catalytic reaction. 相似文献
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Funke SA Otte N Eggert T Bocola M Jaeger KE Thiel W 《Protein engineering, design & selection : PEDS》2005,18(11):509-514
Chiral compounds can be produced efficiently by using biocatalysts. However, wild-type enzymes often do not meet the requirements of a production process, making optimization by rational design or directed evolution necessary. Here, we studied the lipase-catalyzed hydrolysis of the model substrate 1-(2-naphthyl)ethyl acetate both theoretically and experimentally. We found that a computational equivalent of alanine scanning mutagenesis based on QM/MM methodology can be applied to identify amino acid positions important for the activity of the enzyme. The theoretical results are consistent with concomitant experimental work using complete saturation mutagenesis and high-throughput screening of the target biocatalyst, a lipase from Bacillus subtilis. Both QM/MM-based calculations and molecular biology experiments identify histidine 76 as a residue that strongly affects the catalytic activity. The experiments demonstrate its important influence on enantioselectivity. 相似文献