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Steffen Wolf Benedikt Sohmen Bjrn Hellenkamp Johann Thurn Gerhard Stock Thorsten Hugel 《Chemical science》2021,12(9):3350
We report on a study that combines advanced fluorescence methods with molecular dynamics (MD) simulations to cover timescales from nanoseconds to milliseconds for a large protein. This allows us to delineate how ATP hydrolysis in a protein causes allosteric changes at a distant protein binding site, using the chaperone Hsp90 as test system. The allosteric process occurs via hierarchical dynamics involving timescales from nano- to milliseconds and length scales from Ångstroms to several nanometers. We find that hydrolysis of one ATP is coupled to a conformational change of Arg380, which in turn passes structural information via the large M-domain α-helix to the whole protein. The resulting structural asymmetry in Hsp90 leads to the collapse of a central folding substrate binding site, causing the formation of a novel collapsed state (closed state B) that we characterise structurally. We presume that similar hierarchical mechanisms are fundamental for information transfer induced by ATP hydrolysis through many other proteins.We report on a study that combines advanced fluorescence methods with molecular dynamics simulations to cover timescales from nanoseconds to milliseconds for a large protein, the chaperone Hsp90. 相似文献
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W. Pilz und Ilse Johann 《Fresenius' Journal of Analytical Chemistry》1965,210(2):113-123
Zusammenfassung Es wird eine neue Methode zur spezifischen Bestimmung von freiem -Naphthol in Gegenwart eines großen Überschusses von -Naphthylesteremulsion beschrieben. Alle notwendigen reaktionskinetischen und optischen Bedingungen werden untersucht. Die Methode eignet sich besonders gut zur enzymatischen Analyse von Fermenten, die -Naphthylester hydrolysieren. Nach genauer Beschreibung von Arbeitsvorschriften werden folgende Anwendungsbeispiele ausgeführt: a) die Bestimmung von freiem -Naphthol in rohen -Naphthylesterpräparaten, b) die Bestimmung von Enzymaktivitäten: Substratvariation im menschlichen Serum, c) Verwendung als Suchreaktion: Ortung der -Naphthylpropionat spaltenden Fermente und Proteine des menschlichen Serums durch biologische Analyse der Eluate nach präparativer Stärkebreielektrophorese. Die Empfindlichkeit der Methode beträgt mindestens 0,1 Mol -Naphthol; die Störanfälligkeit ist minimal.
Meinem verehrten Lehrer Herrn Univ. Prof. Dr. Alois Musil zum 65. Geburtstag gewidmet. 相似文献
Summary A new method is described for the specific determination of free -naphthol in presence of a large excess of emulsions of -naphthol esters. The kinetics of the reaction and all optical conditions are tested. The method is applicable to the determination of esterases which hydrolyse -naphthol esters. The following examples for the use of the method are described: a) determination of free -naphthol in industrial -naphthol ester preparations, b) determination of esterase activities; c) application as finding test: localisation of the -naphthol propionic acid ester hydrolysing esterases and proteins of human serum by biological analysis of the eluates after preparative starch electrophoresis. The sensitivity is at least 0.1 Mol of -naphthol; the security is optimal.
Meinem verehrten Lehrer Herrn Univ. Prof. Dr. Alois Musil zum 65. Geburtstag gewidmet. 相似文献
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Robert J. Houben Hans-Gerd M. Janssen Piet A. Leclercq Jacques A. Rijks Carel A. Cramers 《Journal of separation science》1990,13(10):669-673
A simple and versatile system is described for the on-line coupling of SFE to capillary GC. The interfacing consists of a programmed temperature vaporizer (PTV) injector. With this injector it is possible to combine solute trapping, elimination of a high flow of extraction fluid, and quantitative transfer of solutes to the seperation column. The problems caused by impurities in the extraction fluid in on-line SFE-GC are discussed. Simple methods are described for the purification of commercially available carbon dioxide. The trapping efficiency of the PTV injector is studied. Applications of the SFE-PTV-GC system are given for the analysis of polymer anti-degradants, polar compounds, and samples with environmental relevance. 相似文献
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The emission of NOx from coal-fired boilers can be limited by means of the selective catalytic reduction of NOx with ammonia. The amounts of unreacted ammonia downstream should be low to avoid processing and environmental problems. Continuous measurement of the ammonia in the flue gas is needed. The determination of ammonia and flue gas sampling techniques are discussed. Measurements of ammonia in exhausts of a laboratory reactor and of a pilot plant for the selective catalytic reduction of NOx with ammonia are presented. Ammonia was determined by mass spectrometry and chemiluminescence in the gas phase, and by spectrophotometric (Nessler and Berthollet reactions) or potentiometry in aqueous solution, in low (<5 μl l?1) and high (<1000 μl l?1) concentration ranges. 相似文献
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Helmut Simon Johann Bader Helmut Günther Stefan Neumann Jordanes Thanos 《Angewandte Chemie (International ed. in English)》1985,24(7):539-553
It has been known for many decades that chiral compounds can be obtained by stereospecific biocatalytic reduction. Further significant methodological developments in this field have, however, only been made during the past ten years; they include the application of previously unused microorganisms and electron donors, the discovery of additional substrates for the known reductases, the development of methods for regenerating reduced pyridine nucleotides, and the discovery of new reductases which were sought for specific preparative purposes. Many chiral compounds can now be synthesized by microbial hydrogenation using H2 and hydrogenase-containing microorganisms as well as by electromicrobial or electroenzymatic reduction. In the two latter methods, anaerobic or aerobic organisms are supplied with electrons from electrochemically reduced, artificial mediators, e.g., methyl viologen. Reductases that do not require pyridine nucleotides and can accept electrons directly from reduced viologens are especially useful. Two examples of this type of enzyme are described which are of preparative interest. Many cells contain methyl viologen-dependent NAD(P) reductases, a large number of which have still not been characterized. A productivity number is proposed which allows different methods of bioconversion with microorganisms to be compared. The productivity numbers of compounds synthesized by the methods described in this review are often 10- to 100-fold higher than those of substances obtained by conventional techniques. 相似文献
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