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991.
Nonribosomal Peptides Produced by Minimal and Engineered Synthetases with Terminal Reductase Domains
Andreas Tietze Dr. Yan-Ni Shi Dr. Max Kronenwerth Prof. Dr. Helge B. Bode 《Chembiochem : a European journal of chemical biology》2020,21(19):2750-2754
Nonribosomal peptide synthetases (NRPSs) use terminal reductase domains for 2-electron reduction of the enzyme-bound thioester releasing the generated peptides as C-terminal aldehydes. Herein, we reveal the biosynthesis of a pyrazine that originates from an aldehyde-generating minimal NRPS termed ATRed in entomopathogenic Xenorhabdus indica. Reductase domains were also investigated in terms of NRPS engineering and, although no general applicable approach was deduced, we show that they can indeed be used for the production of similar natural and unnatural pyrazinones. 相似文献
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Dr. Barbara Wienen-Schmidt Matthias Oebbeke Dr. Khang Ngo Prof. Dr. Andreas Heine Prof. Dr. Gerhard Klebe 《ChemMedChem》2021,16(1):292-300
In lead optimization, protein crystallography is an indispensable tool to analyze drug binding. Binding modes and non-covalent interaction inventories are essential to design follow-up synthesis candidates. Two protocols are commonly applied to produce protein–ligand complexes: cocrystallization and soaking. Because of its time and cost effectiveness, soaking is the more popular method. Taking eight ligand hinge binders of protein kinase A, we demonstrate that cocrystallization is superior. Particularly for flexible proteins, such as kinases, and larger ligands cocrystallization captures more reliable the correct binding pose and induced protein adaptations. The geometrical discrepancies between soaking and cocrystallization appear smaller for fragment-sized ligands. For larger flexible ligands that trigger conformational changes of the protein, soaking can be misleading and underestimates the number of possible polar interactions due to inadequate, highly impaired positions of protein amino-acid side and main chain atoms. Thus, if applicable cocrystallization should be the gold standard to study protein–ligand complexes. 相似文献
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Concrete spalling due to fire exposure is often defined as the sudden detachment of fragments from a concrete surface. It can be quantified by various parameters of which weight loss and spalling depth are the most common ones. The risk of spalling is influenced by many factors such as concrete composition, heating rate and applied testing methods. A reduced scale testing method should be developed to analyse the spalling behaviour and to understand its effectiveness in more detail. As a subsection of this development, this study aimed to analyse the effect of different‐sized, circular heated areas in semi full‐scale fire tests. Therefore, vermiculite slabs with varying cut‐outs in their centre were placed between a specimen made of a spalling‐sensitive concrete and the combustion chamber. The combustion chamber was heated following a standard fire curve. Our experimental results show that the thermal expansion inside of equal‐sized specimens is strongly dependent on the size of the heated area. In addition, this area also affects thermal stresses. They decrease as a result of lower temperature gradients for tests with smaller unheated boundary areas. Apart from this, the analysis of fragments shows no correlation between their relative volume distribution and the heated area. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
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Occupants’ satisfaction had been researched independently related to thermal and visual stimuli for many decades showing among others the influence of self-perceived control. Few studies revealed interactions between thermal and visual stimuli affecting occupant satisfaction. In addition, studies including interactions between thermal and visual stimuli are lacking different control scenarios. This study focused on the effects of thermal and visual factors, their interaction, seasonal influences, and the degree of self-perceived control on overall, thermal, and visual satisfaction. A repeated-measures laboratory study with 61 participants running over two years and a total of 986 participant sessions was conducted. Mixed model analyses with overall satisfaction as outcome variable revealed that thermal satisfaction and visual satisfaction are the most important predictors for overall satisfaction with the indoor environment. Self-perceived thermal control served as moderator between thermal satisfaction and overall satisfaction. Season had slight influence on overall satisfaction. Random effects explained the highest amount of variance, indicating that intra- and interindividual differences in the ratings of satisfaction are more prevalent than study condition. Future building design and operation plans aiming at a high level of occupant satisfaction should consider personal control opportunities and take into account the moderating effect of control opportunities in multimodal interactions. 相似文献
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Giuseppe Schiavone Florian Fallegger Xiaoyang Kang Beatrice Barra Nicolas Vachicouras Evgenia Roussinova Ivan Furfaro Sébastien Jiguet Ismael Seáñez Simon Borgognon Andreas Rowald Qin Li Chuan Qin Erwan Bézard Jocelyne Bloch Grégoire Courtine Marco Capogrosso Stéphanie P. Lacour 《Advanced materials (Deerfield Beach, Fla.)》2020,32(17):1906512
The convergence of materials science, electronics, and biology, namely bioelectronic interfaces, leads novel and precise communication with biological tissue, particularly with the nervous system. However, the translation of lab-based innovation toward clinical use calls for further advances in materials, manufacturing and characterization paradigms, and design rules. Herein, a translational framework engineered to accelerate the deployment of microfabricated interfaces for translational research is proposed and applied to the soft neurotechnology called electronic dura mater, e-dura. Anatomy, implant function, and surgical procedure guide the system design. A high-yield, silicone-on-silicon wafer process is developed to ensure reproducible characteristics of the electrodes. A biomimetic multimodal platform that replicates surgical insertion in an anatomy-based model applies physiological movement, emulates therapeutic use of the electrodes, and enables advanced validation and rapid optimization in vitro of the implants. Functionality of scaled e-dura is confirmed in nonhuman primates, where epidural neuromodulation of the spinal cord activates selective groups of muscles in the upper limbs with unmet precision. Performance stability is controlled over 6 weeks in vivo. The synergistic steps of design, fabrication, and biomimetic in vitro validation and in vivo evaluation in translational animal models are of general applicability and answer needs in multiple bioelectronic designs and medical technologies. 相似文献
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Carina A. Sommer Dr. Andreas Eichinger Prof. Dr. Arne Skerra 《Chembiochem : a European journal of chemical biology》2020,21(4):469-472
Boronic acids have long been known to form cyclic diesters with cis-diol compounds, including many carbohydrates. This phenomenon was previously exploited to create an artificial lectin by incorporating p-borono-l -phenylalanine (Bpa) into the ligand pocket of an engineered lipocalin, resulting in a so-called Borocalin. Here we describe the X-ray analysis of its covalent complex with 4-nitrocatechol as a high-affinity model ligand. As expected, the crystal structure reveals the formation of a cyclic diester between the biosynthetic boronate side chain and the two ortho-hydroxy substituents of the benzene ring. Interestingly, the boron also has a hydroxide ion associated, despite an only moderately basic pH 8.5 in the crystallization buffer. The complex is stabilized by a polar contact to the side chain of Asn134 within the ligand pocket, thus validating the functional design of the Borocalin as an artificial sugar-binding protein. Our structural analysis demonstrates how a boronate can form a thermodynamically stable diester with a vicinal diol in a tetrahedral configuration in aqueous solution near physiological pH. Moreover, our data provide a basis for the further engineering of the Borocalin with the goal of specific recognition of biologically relevant glycans. 相似文献