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排序方式: 共有257条查询结果,搜索用时 46 毫秒
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Paula Bracco Hein J. Wijma Bastian Nicolai Jhon Alexander Rodriguez Buitrago Thomas Klünemann Agustina Vila Patrick Schrepfer Wulf Blankenfeldt Dick B. Janssen Prof. Dr. Anett Schallmey 《Chembiochem : a European journal of chemical biology》2021,22(6):1099-1110
CYP154C5 from Nocardia farcinica is a P450 monooxygenase able to hydroxylate a range of steroids with high regio- and stereoselectivity at the 16α-position. Using protein engineering and substrate modifications based on the crystal structure of CYP154C5, an altered regioselectivity of the enzyme in steroid hydroxylation had been achieved. Thus, conversion of progesterone by mutant CYP154C5 F92A resulted in formation of the corresponding 21-hydroxylated product 11-deoxycorticosterone in addition to 16α-hydroxylation. Using MD simulation, this altered regioselectivity appeared to result from an alternative binding mode of the steroid in the active site of mutant F92A. MD simulation further suggested that the entrance of water to the active site caused higher uncoupling in this mutant. Moreover, exclusive 15α-hydroxylation was observed for wild-type CYP154C5 in the conversion of 5α-androstan-3-one, lacking an oxy-functional group at C17. Overall, our data give valuable insight into the structure–function relationship of this cytochrome P450 monooxygenase for steroid hydroxylation. 相似文献
3.
Benedikt Ley Corinna Ogonowski Jan Hess Tim Reichling Lin Wan Volker Wulf 《Behaviour & Information Technology》2014,33(8):815-828
Technological infrastructure at home is changing continuously and is becoming increasingly interconnected. Media devices, including the TV set, provide access to the Internet and offer manifold opportunities to consume media on demand. Additionally, personal devices, such as smartphones, also enable flexible consumption and sharing of media. Questions about how these technologies change the user's media usage and how these changes affect the social structure of a household, however, remain largely unanswered. In order to gain insight into the adoption of new technologies into daily routines, we explored these changes in respect of people's media usage in a qualitative long-term Living Lab study. We will present findings regarding personal routines, flexible integration of new devices into existing practices, influences on households as social systems and related issues in device access and collective use. We will highlight potentials and conflicts regarding device shifts and roles; restrictions in device access; social influences in the living room; and individual changes in media consumption. 相似文献
4.
Rafael Prikladnicki Alexander Boden Gabriela Avram Cleidson R. B. de Souza Volker Wulf 《Empirical Software Engineering》2014,19(4):822-856
Global Software Engineering has become a standard in today’s software industry. Research in distributed software development poses severe challenges that are due to the spatial and temporal distribution of the actors, as well as to language, intercultural and organizational aspects. These challenges occur in addition to “traditional” challenges of the domain itself in large-scale software projects, like coordination and communication issues, requirements volatily, lack of domain knowledge, among others. While several authors have reported empirical studies of global software development projects, the methodological difficulties and challenges of this type of studies have not been sufficiently discussed. In this paper, we share our experiences of collecting and analysing qualitative data in the context of Global Software Engineering projects. We discuss strategies for gaining access to field sites, building trust and documenting distributed and complex work practices in the context of several research projects we have conducted in the past 9 years. The experiences described in this paper illustrate the need to deal with fundamental problems, such as understanding local languages and different cultures, observing synchronous interaction, or dealing with barriers imposed by political conflicts between the sites. Based on our findings, we discuss some practical implications and strategies that can be used by other researchers and provide some recommendations for future research in methodological aspects of Global Software Engineering. 相似文献
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Kai Schubert Anne Weibert Volker Wulf 《International journal of human-computer studies》2011,69(10):669-678
Located in socially and culturally diverse neighborhoods, we have built a network of intercultural computer clubs, called come_IN. These clubs offer a place to share practices among children and adults of diverse ethnical backgrounds. We show how this initiative ties into the striving for the integration of migrant communities and host society in Germany. In this paper, we analyze how collaborative project work and the use of mobile media and technologies contribute to integration processes in multicultural neighborhoods. Qualitative data gathered from interviews with club participants, participative observation in the computer clubs, as well as the analysis of artifacts created during project work provides the background needed to match local needs and peculiarities with (mobile) technologies. Based on these findings we present two approaches to add to the technological infrastructure: (1) a mesh-network extending the clubs into the neighborhood and (2) a project management tool, which supports projects and stimulates the sharing of ideas among projects. 相似文献
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Structures of the N-Terminal Domain of PqsA in Complex with Anthraniloyl- and 6-Fluoroanthraniloyl-AMP: Substrate Activation in Pseudomonas Quinolone Signal (PQS) Biosynthesis
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Florian Witzgall Wiebke Ewert Prof. Dr. Wulf Blankenfeldt 《Chembiochem : a European journal of chemical biology》2017,18(20):2045-2055
Pseudomonas aeruginosa, a prevalent pathogen in nosocomial infections and a major burden in cystic fibrosis, uses three interconnected quorum-sensing systems to coordinate virulence processes. At variance with other Gram-negative bacteria, one of these systems relies on 2-alkyl-4(1H)-quinolones (Pseudomonas quinolone signal, PQS) and might hence be an attractive target for new anti-infective agents. Here we report crystal structures of the N-terminal domain of anthranilate-CoA ligase PqsA, the first enzyme of PQS biosynthesis, in complex with anthraniloyl-AMP and with 6-fluoroanthraniloyl-AMP (6FABA-AMP) at 1.4 and 1.7 Å resolution. We find that PqsA belongs to an unrecognized subfamily of anthranilate-CoA ligases that recognize the amino group of anthranilate through a water-mediated hydrogen bond. The complex with 6FABA-AMP explains why 6FABA, an inhibitor of PQS biosynthesis, is a good substrate of PqsA. Together, our data might pave a way to new pathoblockers in P. aeruginosa infections. 相似文献
9.
AbstractThe objective of this study was to compare spray-dried and freeze-dried extracts of Sennae fructus regarding the stability of the sennosides. Therefore, the influence of the addition of maltodextrin (MD) not only before the drying processes but also during fluidized bed granulation was investigated. Analysis of the hygroscopicities, the simultaneous TGA-DSC-MS data, and the influence of storage revealed that spray-dried extracts are more stable than freeze-dried extracts. The addition of MD led to an even better stability of the extracts. Lower amounts of MD are sufficient if applied onto the surface of the native extract during fluidized bed granulation. 相似文献
10.
The Alkylquinolone Repertoire of Pseudomonas aeruginosa is Linked to Structural Flexibility of the FabH‐like 2‐Heptyl‐3‐hydroxy‐4(1H)‐quinolone (PQS) Biosynthesis Enzyme PqsBC
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Florian Witzgall Tobias Depke Dr. Michael Hoffmann Dr. Martin Empting Prof. Dr. Mark Brönstrup Prof. Dr. Rolf Müller Prof. Dr. Wulf Blankenfeldt 《Chembiochem : a European journal of chemical biology》2018,19(14):1531-1544
Pseudomonas aeruginosa is a bacterial pathogen that causes life‐threatening infections in immunocompromised patients. It produces a large armory of saturated and mono‐unsaturated 2‐alkyl‐4(1H)‐quinolones (AQs) and AQ N‐oxides (AQNOs) that serve as signaling molecules to control the production of virulence factors and that are involved in membrane vesicle formation and iron chelation; furthermore, they also have, for example, antibiotic properties. It has been shown that the β‐ketoacyl‐acyl‐carrier protein synthase III (FabH)‐like heterodimeric enzyme PqsBC catalyzes the last step in the biosynthesis of the most abundant AQ congener, 2‐heptyl‐4(1H)‐quinolone (HHQ), by condensing octanoyl‐coenzyme A (CoA) with 2‐aminobenzoylacetate (2‐ABA), but the basis for the large number of other AQs/AQNOs produced by P. aeruginosa is not known. Here, we demonstrate that PqsBC uses different medium‐chain acyl‐CoAs to produce various saturated AQs/AQNOs and that it also biosynthesizes mono‐unsaturated congeners. Further, we determined the structures of PqsBC in four different crystal forms at 1.5 to 2.7 Å resolution. Together with a previous report, the data reveal that PqsBC adopts open, intermediate, and closed conformations that alter the shape of the acyl‐binding cavity and explain the promiscuity of PqsBC. The different conformations also allow us to propose a model for structural transitions that accompany the catalytic cycle of PqsBC that might have broader implications for other FabH‐enzymes, for which such structural transitions have been postulated but have never been observed. 相似文献